Chemical compounds

ABSTRACT

Aryl substituted phosphoryl derivatives of the formula                  
 
In which Ar is phenyl, naphthyl, or pyridyl, Y is O or S, X 1  is O, NR 3 , S, CR 3 R 4 , CR 3 W 1  or CW 1 W 2 , X 2  and X 6  are a bond or X 6  is CH 2  and X 2  is O, NR 3 , S, CR 3 R 4 , CR 3 W 1  or CW 1 W 2 , R 3  and R 4  are H, alkyl or phenyl, groups, W 1  and W 2  are heteroatoms, X 3  is alkylene, X 4  is oxygen or CH 2 , X 5  is a bond or CH 2 , Z is O, NR 5 , S, alkyl or phenyl, R 5  is H, alkyl or phenyl, J is H, alkyl, phenyl, or a heterocyclic or polycyclic group, Q is O, NR 6 , S, CR 6 R 7 , CR 6 W 3  or CW 3 W 4 , R 6  and R 7  are H, alkyl or phenyl, and W 3  and W 4  are hetero atoms, T 1  and T 2  are H or CH 2 R 8 , R 8  is H, OH or F, or T 1  and T 2  together are —CH═CH— or —C(R 9 )(R 10 )C(R 11 )(R 12 )—, R 9  is H, halogeno, CN, NH 2 , CO-alkyl, or alkyl, R 10 , R 11 , and R 12  are H, N 3 , halogen, CN, NH 2 , CO-alkyl, or alkyl, and B is a purine or pyrimidine base, have antiviral activity, as for example against HIV. Particularly preferred are thymine and adenine derivatives of amino acid phenoxyphosphoroamidates. A typical embodiment is 2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenyl methoxy alaninyl) phosphoroamidate which can be prepared from phenyl methoxy alaninyl phosphorochloridate and 2′,3′-dideoxy-2′,3′-didehydrothymidine.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of U.S. patent applicationSer. No. 09/382,084, filed Aug. 24, 1999, now U.S. Pat. No. 6,455,513,which in turn is a continuation of U.S. patent application Ser. No.08/913,639, filed Feb. 2, 1998, now abandoned, which is 371 ofapplication No. PCT/GB96/00580, filed Mar. 13, 1996.

The present invention relates to a new class of nucleoside analogues andtheir therapeutic use in the prophylaxis and treatment of viralinfection, for example by human immunodeficiency virus (HIV), which isbelieved to be the aetiological agent in human acquired immunodeficiencysyndrome (AIDS).

There has been much interest in the use of nucleoside analogues asinhibitors of HIV. 2′,3′-dideoxy-2′,3′-didehydrothymidine (d4T) and3′-azido-3′-deoxythymidine (AZT) are both known inhibitors of HIV[Hitchcock et al., Antiviral Chem. Chemother. (1991), 2, 125; Mansuri etal., Antimicrob. Agents Chemother., (1990), 34, 637.]. The inhibition ofHIV by these, and other nucleoside analogues, is conventionally thoughtto depend upon conversion of the nucleoside analogue in vivo to thecorresponding 5′-triphosphate by (host-cell) kinase enzymes. However,this absolute dependence upon (host-cell) kinase-mediated activation canlead to poor activity, the emergence of resistance, and clinicaltoxicity.

In order to reduce the dependence on kinase enzymes the use of maskedphosphate pro-drugs of the bioactive nucleotide forms of severalchemotherapeutic nucleoside analogues has been suggested [McGuigan etal., Nucleic Acids Res., (1989), 17, 6065; McGuigan et al., Ibid.,(1989), 17, 7195; Chawla et al., J. Med. Chem., (1984), 27, 1733;Sergheraert et al., J. Med. Chem. (1993), 36, 826–830.]. In particular,McGuigan et al [J. Med. Chem. 36, 1048–1052 (1993)] have reported thepreparation of aryl ester-phosphoramidate derivatives of AZT. In vitroevaluation of these compounds revealed the compounds to have anti-HIVactivity. However, in “normal” thymidine kinase rich (TK⁺) cells, theactivity of such compounds was at least an order of magnitude less thanthe parent nucleoside AZT. Only in TK-deficient (TK⁻) cells, in whichthe activity of the aryl ester-phosphoramidate derivatives was virtuallymaintained but the activity of AZT was reduced, did the activity of thederivatives exceed that of AZT.

McGuigan et al [Bioorganic & Medical Chemistry Letters, 3, (6),1203–1206 (1993)] have also reported preparation of triester phosphatederivatives of d4T. Again, in vitro evaluation of these compoundsrevealed that whilst the compounds have significant anti-HIV activity,the activity is less than that of the parent nucleoside d4T in TK⁺cells.

Abraham and Wagner (Nucleosides and Nucleotides 13 (9). 1891–1903(1994)) have reported the preparation of nucleoside phosphoramidatediesters and triesters but do not report any biological activity.

The acyclic nucleoside analogue 9(2-phosphonomethoxyethyl) adenine(PMEA), and analogues thereof, have been demonstrated to show activityagainst herpes viruses and retroviruses including HIV (Calio et al.,Antiviral Res., (1994), 23(1), 77–89; Balzarini et al., AIDS, (1991),5(1), 21–28).

To date, the approach of providing masked phosphate pro-drugs has failedto enhance the anti-viral activities of the parent nucleoside analoguessuch as AZT and d4T in TK⁺ cells. Furthermore, the emergence ofresistance to the nucleoside analogues in their bioactive5′-triphosphate form has rendered the reported masked phosphatepro-drugs and their parent nucleoside analogues potentially ineffective.

It has now been found that a particular class of masked nucleosideanalogues are highly potent viral inhibitors in both TK⁻ and TK⁺ cells,and yet retain activity against nucleoside (e.g. d4T)—resistant virus.

According to the present invention there is provided a compound of theformula (1)

wherein

-   -   Ar is an aryl group;    -   Y is oxygen or sulphur;    -   X¹ is selected from O, NR³, S, CR³R⁴, CR³W¹ and CW¹W² where R³        and R⁴ are independently selected from hydrogen, alkyl and aryl        groups; and W¹ and W² are heteroatoms;    -   X²–X⁶ may be absent; or X⁶ is CH₂ and X² is selected        (independently of X¹) from O, NR³, S, CR³R⁴, CR³W¹ and CW¹W²        where R³ and R⁴ are independently selected from hydrogen, alkyl        and aryl groups; and W¹ and W² are heteroatoms;    -   X³ is a C₁₋₆ alkyl group;    -   X⁴ is oxygen or CH₂;    -   X⁵ may be absent or is CH₂;    -   Z is selected from O, NR⁵, S, alkyl and aryl groups, where R⁵ is        selected from hydrogen, alkyl and aryl groups;    -   J is selected from hydrogen, alkyl, aryl, heterocyclic and        polycyclic groups;    -   Q is selected from O, NR⁶, S, CR⁶R⁷, CR⁶W³ and CW³W⁴, where R⁶        and R⁷ are independently selected from hydrogen, alkyl and aryl        groups; and W³ and W⁴ are heteroatoms;    -   T¹ and T² are independently selected from hydrogen and CH₂R⁸,        where R⁸ is selected from H, OH and F; or T¹ and T² are linked        together and together are selected from the groups        -   where R⁹ is selected from H, halogen, CN, NH₂, CO-alkyl and            alkyl; and R¹⁰, R¹¹, R¹² are independently selected from H,            N₃, halogen, CN, NH₂, CO-alkyl and alkyl;    -   B is a purine or pyrimidine base;        or a pharmaceutically acceptable derivative or metabolite        thereof.

The compounds of the present invention are potent anti-viral agents. Inparticular, they are highly active against HIV in both TK⁻ and TK⁺cells. Particularly surprising is the activity of the compounds of thepresent invention against nucleoside-resistant HIV. These observationsindicate that the activity of these compounds is not wholly dependentupon the conventional mode of action (requiring hydrolysis of thephosphate aryl ester and P—X¹ bonds followed by kinase-dependentconversion to the 5′-triphosphate derivative), but arises from anentirely different mode of action. The experimental data presentedherein indicates that the compounds and metabolites of the presentinvention are directly acting as reverse transcriptase (RT) inhibitorsvia a previously unrecognised metabolic pathway and mechanism of action.

Reference in the present specification to an alkyl group means abranched or unbranched, cyclic or acyclic, saturated or unsaturated(e.g. alkenyl or alkynyl) hydrocarbyl radical. Where cyclic, the alkylgroup is preferably C₃ to C₁₂, more preferably C₅ to C₁₀, morepreferably C₅ to C₇. Where acyclic, the alkyl group is preferably C₁ toC₁₆, more preferably C₁ to C₆, more preferably methyl. Reference in thepresent specification to alkoxy and aryloxy groups means alkyl-O— andaryl-O— groups, respectively. Reference to alkoyl and aryloyl groupsmeans alkyl-CO— and aryl-CO—, respectively.

Reference in the present specification to an aryl group means anaromatic group, such as phenyl or naphthyl, or a heteroaromatic groupcontaining one or more, preferably one, heteroatom, such as pyridyl,pyrrolyl, furanyl and thiophenyl. Preferably, the aryl group comprisesphenyl or substituted phenyl.

The alkyl and aryl groups may be substituted or unsubstituted,preferably unsubstituted. Where substituted, there will generally be 1to 3 substituents present, preferably 1 substituent. Substituents mayinclude halogen atoms and halomethyl groups such as CF3 and CC 13;oxygen containing groups such as oxo, hydroxy, carboxy, carboxyalkyl,alkoxy, alkoyl, alkoyloxy, aryloxy, aryloyl and aryloyloxy; nitrogencontaining groups such as amino, alkylamino, dialkylamino, cyano, azideand nitro; sulphur containing groups such as thiol, alkylthiol,sulphonyl and sulphoxide; heterocyclic groups which may themselves besubstituted; alkyl groups, which may themselves be substituted; and arylgroups, which may themselves be substituted, such as phenyl andsubstituted phenyl. Alkyl includes substituted and unsubstituted benzyl.Reference in the present specification to heterocyclic groups meansgroups containing one or more, pyrrolyl, imidazolyl, pyrazolyl,thiazolyl, isothiazolyl, oxazolyl, pyrrolidinyl, pyrrolinyl,imidazolidinyl, imidazolinyl, pyrazolidinyl, tetrahydrofuranyl, pyranyl,pyronyl, pyridyl, pyrazinyl, pyridazinyl, piperidyl, piperazinyl,morpholinyl, benzofuranyl, isobenzofuryl, indolyl, oxyindolyl,isoindolyl, indazolyl, indolinyl, 7-azaindolyl, isoindazolyl,benzopyranyl, coumarinyl, isocoumarinyl, quinolyl, isoquinolyl,naphthridinyl, cinnolinyl, quinazolinyl, pyridopyridyl, benzoxazinyl,quinoxadinyl, chromenyl, chromanyl, isochrcmanyl and carbolinyl.

References in the present specification to polycyclic groups means agroup comprising two or more non-aromatic carbcyclic or heterocyclicrings which may themselves be substituted.

Reference in the present specification to halogen means a fluorine,chlorine, bromine or iodine radical, preferably fluorine or chlorineradical.

The group Ar comprises a substituted or unsubstituted aryl group,wherein the term “aryl group” and the possible substitution of saidgroup is as defined above. Preferably, Ar is a substituted orunsubstituted phenyl group. Particularly preferred substituents areelection withdrawing groups such as halogen (preferably chlorine orfluorine), trihalomethyl (preferably trifluoromethyl), cyano and nitrogroups. Preferably, Ar is phenyl, 3,5-dichloro-phenyl,p-trifluoromethyl-phenyl, p-cyano-phenyl, or p-nitro-phenyl.

Y may be oxygen or sulphur. Preferably, Y is oxygen.

X¹ is from O, NR³, S, CR³R⁴, CR³W¹ and CW¹W² where R³ and R⁴ areindependently selected from hydrogen, alkyl and aryl groups; and W¹ andW² are heteroatoms. Preferably, X¹ is selected from O, S and NR³.Preferably, X¹ is NR³. When present, R³ is preferably H. When present,W¹ and W² may independently comprise any heteroatom such as a halogen,preferably fluorine.

X²–X⁶ may be absent; or X⁶ is CH₂ and X² is selected (independently ofX¹) from O, NR³, S, CR³R⁴, CR³W¹ and CW¹W² where R³ and R⁴ areindependently selected from H, alkyl and aryl groups; and W¹ and W² areheteroatoms. When present, X² is preferably oxygen. When present, R³ ispreferably H. When present W¹ and W² may independently comprise anyheteroatom such as halogen, preferably fluorine.

X⁴ is oxygen or CH₂. Preferably, X⁴ is oxygen.

X⁵ may be absent or is CH₂.

Z may comprise O, NR⁵, S, alkyl or aryl groups, where R⁵ is selectedfrom H, alkyl and aryl groups. Preferably, Z is O or NR⁵. Preferably, R⁵is hydrogen. Most preferably, Z is oxygen.

J is selected from hydrogen, alkyl, aryl, heterocyclic and polycyclicgroups. Preferably, J is a substituted or unsubstituted alkyl group.Preferably, J is a substituted or unsubstituted C₁₋₆ alkyl group,preferably a benzyl or methyl group.

X³ is a C₁₋₆ alkyl group. X³ may be a C₁₋₆ substituted or unsubstituted,branched or unbranched, methylene chain. Preferably, X³ is a group CR¹R²where R¹and R² are independently selected from hydrogen, alkyl and arylgroups. Preferably, at least one of R¹ and R² is hydrogen. It will beappreciated that if R¹and R² are different, the carbon atom to whichthey are bonded is an asymmetric centre. The stereochemistry at thissite may be R or S or mixed. When one of R³ and R⁴ is hydrogen, thestereochemistry is preferably S.

Q is selected from O, NR⁶, S, CR⁶R⁷, CR⁶W³ and CW³W⁴, where R⁶ and R⁷are independently selected from hydrogen, alkyl and aryl groups; and W²and W³ are heteroactoms such as halogen atoms, preferably fluorine.Preferably, Q is O, S, CH₂ or CF₂. Most preferably, Q is oxygen.

T¹ and T² are independently selected from hydrogen and CH₂R⁸ where R⁸ isselected from H, OH and F; or T² and T² are linked together and togetherare selected from the groups:

where R⁹ is selected from H, halogen, CN, NH₂, CO-alkyl, and alkyl,preferably R⁹ is H or F; and R¹⁰, R¹¹, and R¹² are independentlyselected from H, N₃, halogen, CN, NH₂, CO-alkyl, and alkyl, preferablyR¹⁰, R¹¹ and R¹² are independently selected from H, F and N₃. It will beappreciated that R⁹ corresponds to the 3′—α position and R¹⁰ correspondsto the 3′—β position. Preferably, T¹ and T² are linked together andtogether form the group:

B comprises a purine or pyrimidine base, such as adenine thymine,uracil, cytosine or guanine and derivatives thereof. Derivatives thereofinclude substituted purine or pyrimidine bases wherein the substituentsare as defined above. Examples of substituted bases include5-substituted pyrimidine. Preferably, B is adenine or thymine.

Preferably, the present invention provides a compound of formula (2)

wherein Ar, R¹, J, X², X⁵, X⁶, Q, T¹, T² and B are as defined above; ora pharmaceutically acceptable derivative or metabolite thereof.

It will be appreciated that the group —NH—CHR¹—CO₂J corresponds to acarboxy-protected α-amino acid. Preferably, the group R¹ corresponds tothe side chain of a naturally occurring amino acid such as Alanine,Arginine, Asparagine, Aspartic Acid, Cysteine, Cystine, Glycine,Glutamic Acid, Glutamine, Histidine, Isoleucine, Leucine, Lysine,Methionine, Phenylalanine, Proline, Serine, Threonine, Tryptophan,Tyrosine, Valine. Preferably, R¹ is Me or PhCH₂ corresponding to theside chain of alanine or phenylalanine, respectively. Preferably, thestereochemistry at the asymmetric center —CHR¹— corresponds to anL-amino acid.

According to one preferred embodiment, the present invention provides acompound of formula (3):

wherein Ar, Y, X¹, X², X³, X⁴, Z, Q and B are as defined above.

More preferably, the invention provides a compound, according to formula(3), of formula (4):

wherein Ar, R¹ and J are as defined above; or a pharmaceuticallyacceptable derivative or metabolite thereof. Preferably, the inventionprovides a compound of formula (4) in which Ar, R¹ and J are defined inaccordance with Table 1.

TABLE 1 Compound Reference Ar R¹ J 323 4-EtPh Me Me 324 Ph Me Me 3274-FPh Me Me 526 3-CF₃Ph Me Me 546 3,5-Cl₂Ph Me Me 730 Ph Me Bzl 7762,4-Br₂Ph Me Me 779 F₅Ph Me Me 862 Ph Me Hexyl 863 Ph Bzl Me 864 PhCH₂iPr Me 865 Ph iPr Me 866 Ph H Me 867 Ph [CH₂]₂SMe Me 868 2,4Br₂Ph MeBzl 877 Ph Bzl Bzl 878 Ph Bzl tBu 892 Ph Me Cyclohexyl 893 Ph Me tBu1078 Ph CH₂CO₂H Me 1214 Ph CH₂CH₂CH₂NHC[NH₂]NH Me 1218 Ph Me n-Pent 1219Ph Me neo-Pent 1226 Ph Me 1-Napthyl 1227 Ph Me 2-Napthyl

According to a further preferred embodiment, the present inventionprovides a compound of formula (5)

wherein Ar, Y, X¹, X², X³, X⁴, Z, J, R⁹, R¹⁰, R¹¹, R¹², Q and B asdefined above.

More preferably, the invention provides a compound, according to formula(5), of the formula (6):

wherein Ar, R¹, J, R⁹, R¹⁰, R¹¹, R¹² and B are as defined above.

According to a further preferred embodiment, the present inventionprovides a compound of formula (7):

wherein Ar, Y, X¹, X³, X⁴, Z, J, Q and B are as defined above and T¹ andT² are independently selected from H and CH₂R⁸ wherein R⁸ is as definedabove. Preferably, B is a purine base. More preferably, B is adenine.Preferably, T¹ is hydrogen. Preferably, T² is CH₂R⁸. These compounds areanalogues of the acyclic nucleoside analogue9-(2-phosphonylmethoxyethyl) adenine (PMEA), which has been demonstratedto show activity against herpes viruses and retroviruses (Calio et al.,Antiviral Res., (1994), 23(1), 77–89; Balzarini et al., AIDS, (1991),5(1), 21–28).

More preferably, the invention provides a compound, according to formula(7), of formula (8):

wherein Ar, R¹, J, T¹, T² and B are as defined above.

It is a feature of the aryl ester phosphate compounds (1) of the presentinvention that they exhibit significantly enhanced anti-viral efficacy,in both in vitro and in vivo tests, in comparison to their correspondingnucleoside analogue (9)

In addition, the compounds of the present invention exhibitsignificantly reduced toxicity in comparison to their correspondinganalogue (9).

The compounds of the present invention thus exhibit a greatly enhancedselectivity index (ratio of CC₅₀ (toxicity):EC₅₀ (activity)) incomparison to their corresponding nucleoside analogue.

Experiments with radiolabelled compounds of the present invention haveshown that the compounds give enhanced intracellular levels ofnucleoside 5′-triphosphate, the enhancement being particularlysignificant in TK⁻ cells. Thus, the compounds of the present inventionmay act in part by the known metabolic pathway.

However, it has been found that the compounds of the present inventionshow surprising activity against nucleoside resistant strains of HIV.This indicates that the compounds of the present invention are alsoacting by a pathway independent of a 5′-triphosphate metabolite.

It has been demonstrated that the compounds of the present inventionlead to intracellular generation of high levels of a metabolite (10).

Metabolite (10) may also be prepared by treatment of the correspondingcompound according to formula (1) with hog liver esterase. Moreover, ithas been shown that compounds of formula (10) are direct inhibitors ofreverse transcriptase from HIV.

According to a further aspect of the present invention there is provideda compound of formula (10)

wherein Ar, Y, X¹, X², X³, X⁴, X⁶, T¹, T², Q, X⁵ and B are as definedabove, or a pharmaceutically acceptable derivative or metabolitethereof.

The intracellular generation of anti-viral metabolites such as (10) isan important feature of the invention for several reasons. Firstly, thedirect activity of (10) on RT removes the necessity for furthernucleotide-kinase mediated phosphorylation, which may be slow in manycases. In cases where the nucleoside monophosphate is not a substratefor host nucleotide kinases, activation will be poor and anti-viralefficacy low, even if the triphosphate is an excellent RT inhibitor. Insuch cases, the generation of metabolites such as (10) may lead to avery significant enhancement in antiviral action. Such compounds may beacting directly in their own right or via a rearrangement, decompositionor disproportionation product or via a contaminant. Moreover, thestructure of metabolites such as (10) may be further designed tooptimise binding to the known structure of RT, and such modifiedmetabolites could be delivered intracellularly using technology hereindescribed, to further enhance the anti-viral effect.

By “a pharmaceutically acceptable derivative” is meant anypharmaceutically acceptable salt, ester or salt of such ester or anyother compound which upon administration to a recipient is capable ofproviding (directly or indirectly) a compound of formula (1) or (10). By“pharmaceutically acceptable metabolite” is meant a metabolite orresidue of a compound of formula (1) or (10) which gives rise to anucleoside-resistance independent or nucleoside 5′-triphosphateindependent mode of reverse transcriptase inhibition exhibited by thecompounds of formula (1) or (10).

According to a further aspect of the present invention there is provideda compound according to the present invention for use in a method oftreatment, preferably in the prophylaxis or treatment of viralinfection.

According to a further aspect of the present invention there is provideduse of a compound according to the present invention in the manufactureof a medicament for the prophylaxis or treatment of viral infection.

According to a further aspect of the present invention there is provideda method of prophylaxis or treatment of viral infection comprisingadministration to a patient in need of such treatment an effective doseof a compound according to the present invention.

The viral infection may comprise any viral infection such as HIV andherpes virus, including HSV 1 and HSV 2, CMV, VZV, EBV, HAV, HBV, HCV,HDV, papilloma, rabies and influenza.

Preferably, the viral infection comprises HIV infection, more preferablyHIV-I or HIV-II. It is a feature of the present invention that thecompounds exhibit good activity against both HIV-I and HIV-II.

According to a further aspect of the present invention there is provideduse of a compound of the present invention in the manufacture of amedicament for use in the inhibition of a reverse transcriptase by anucleoside-resistance independent or nucleoside 5′-triphosphateindependent mode of action.

According to a further aspect of the present invention there is provideda pharmaceutical composition comprising a compound of the presentinvention in combination with a pharmaceutically acceptable excipient.

According to a further aspect of the present invention there is provideda method of preparing a pharmaceutical composition comprising the stepof continuing a compound of of the present invention with apharmaceutically acceptable excipient.

The medicaments employed in the present invention can be administered byoral or parenteral routes, including intravenous, intramuscular,intraperitoneal, subcutaneous, transdermal, airway (aerosol), rectal,vaginal and topical (including buccal and sublingual) administration.

For oral administration, the compounds of the invention will generallybe provided in the form of tablets or capsules, as a powder or granules,or as an aqueous solution or suspension.

Tablets for oral use may include the active ingredient mixed withpharmaceutically acceptable excipients such as inert diluents,disintegrating agents, binding agents, lubricating agents, sweeteningagents, flavouring agents, colouring agents and preservatives. Suitableinert diluents include sodium and calcium carbonate, sodium and calciumphosphate, and lactose, while corn starch and alginic acid are suitabledisintegrating agents. Binding agents may include starch and gelatin,while the lubricating agent, if present, will generally be magnesiumstearate, stearic acid or talc. If desired, the tablets may be coatedwith a material such as glyceryl monostearate or glyceryl distearate, todelay absorption in the gastrointestinal tract.

Capsules for oral use include hard gelatin capsules in which the activeingredient is mixed with a solid diluent, and soft gelatin capsuleswherein the active ingredient is mixed with water or an oil such aspeanut oil, liquid paraffin or olive oil.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

For intramuscular, intraperitoneal, subcutaneous and intravenous use,the compounds of the invention will generally be provided in sterileaqueous solutions or suspensions, buffered to an appropriate pH andisotonicity. Suitable aqueous vehicles include Ringer's solution andisotonic sodium chloride. Aqueous suspensions according to the inventionmay include suspending agents such as cellulose derivatives, sodiumalginate, polyvinyl-pyrrolidone and gum tragacanth, and a wetting agentsuch as lecithin. Suitable preservatives for aqueous suspensions includeethyl and n-propyl p-hydroxybenzoate.

The compounds of the invention may also be presented as liposomeformulations.

In general a suitable dose will be in the range of 0.1 to 300 mg perkilogram body weight of the recipient per day, preferably in the rangeof 6 to 150 mg per kilogram body weight per day and most preferably inthe range 15 to 100 mg per kilogram body weight per day. The desireddose is preferably presented as two, three, four, five or six or moresub-doses administered at appropriate intervals throughout the day.These sub-doses may be administered in unit dosage forms, for example,containing 10 to 1500 mg, preferably 20 to 1000 mg, and most preferably50 to 700 mg of active ingredient per unit dosage form.

According to a further aspect of the present invention there is provideda process for the preparation of a compound according to the presentinvention, the process comprising reaction of a compound of formula (11)

with a compound of formula (12)

The reaction may be carried out in the tetrahydrofuran in the presenceof N-methylimidazole.

Alternatively, the compounds of the present invention may be prepared byreaction of a compound of formula (13) or a suitable derivative thereof

with ArOH and a compound of formula (14) or suitable derivatives thereof

The invention will now be described with reference to the followingFigures and Examples. It will be appreciated that what follows is by wayof example only and that modifications to detail may be made whilststill falling within the scope of the invention.

FIG. 1 illustrates the in vivo “activity against virus induced tumourformation” consistent with FIG. 1 and page 60 of the description of d4T(comparative) and aryl ester phosphoramidate compound 324 in MSVinfected mice. Drug doses are 50[low ] or 200 [high] mg/kg/day giveni.p. for 4 days starting 1 hour before MSV inoculation.

Experimental

All experiments involving water sensitive compounds were conducted underscrupulously dry conditions. Tetrahydofuran was dried by heating underreflux over sodium and benzophenone followed by distallation and storageover active sieves. N-methylimidazole was purified by distillation.Nucleosides were dried at elevated temperature in vacuo over P₂O₅.Proton, carbon and phosphorus Nuclear Magnetic Resonance (¹H, ¹³C, ³¹Pnmr) spectra were recorded on a Bruker Avance DPX spectrometer operatingat 300 MHz, 75.5 MHz, and 121.5 MHz respectively. All nmr spectra wererecorded in CDCl₃ at room temperature (20° C. +/−3° C.). ¹H and ¹³Cchemical shifts are quoted in parts per million downfield fromtetramethylsilane. J values refer to coupling constants and signalsplitting patterns are described as singlet (s), broad singlet (bs),doublet (d), triplet (t), quartet (q), multiplet (m) or combinationsthereof. ³¹P chemical shifts are quoted in parts per million relative toan external phosphoric acid standard. Many NMR peaks were further splitdue to the presence of diastereoisomers at the [chiral] phosphatecentre. Chromatography refers to flash column chromatography and wascarried out using Merck silica gel 60H (40–60 m, 230–400 mesh) asstationary phase. Thin layer chromatography was performed using AlugramSIL G/UV₂₅₄ aluminium backed silica gel plates.

Mass spectra were recorded by the fast atom bombardment (FAB) mode on aVG 70–250 spectrometer. HPLC data was recorded using an ACS quaternarysystem with an ODS5 column and an eluent of water/acetonitrile, with 82%water 0–10 mm, and then a linear gradient to 20% water at 30 min, with aflow rate of 2 mL/min and detection by UV at 265 nm.

The test compounds were isolated as mixtures of diastereoisomers, withthis isomerism arising from mixed stereochemistry at the phosphatecentre. The resulting oils did not give useful microanalytical data butwere found to be pure by high-field multinuclear NMR spectroscopy andrigorous HPLC analysis.

Preparation of Compounds

The compounds of the present invention were prepared according to thefollowing general procedures.

Preparation of Aryl Phosphorodichloridates (General Procedure)

A solution of the appropriate phenol (30.4 mmol) and triethylamine (4.25ml, 30.5 mmol) in dry CH₂Cl₂ (25 ml) was added to a solution of freshlydistilled POCl₃ (10 ml, 107 mmol) in CH₂Cl₂ (30 ml) at −50° and themixture allowed to stir at ambient temperature overnight. The reactionmixture was filtered and the filtrate evaporated. Ether (20 ml) wasadded and precipitate filtered again. After evaporation the residue wasdistilled if possible.

Phenyl N-methylalaninyl Phosphorochloridate

A solution of triethylamine (1 ml- 7.17 mmol) in 15 ml of dry CH₂Cl₂ wasadded dropwise to a mixture of phenyl phosphorodichloridate (757.4 mg,3.59 mmol) and L-alanine methyl ester hydrochloride (500 mg, 3.58 mmol)in 50 ml of dry CH₂Cl₂ at −80° C. in one hour. The mixture was thenstirred vigorously at −50° C. during five hours and CH₂Cl₂ evaporated.25 ml of dry ether was added and precipitate filtered off undernitrogen. Evaporation of ether gave a colourless oil which was usedwithout further purification for the next step.

Preparation of Aryl Phosphates of Nucleoside Analogues (GeneralProcedure)

Phenyl N-methylalaninyl phosphorochoridate (250 mg, 0.9 mmol, 2.0equivs) was added to a stirred solution of nucleoside analogue 0.45mmol) and N-methylimidazole (0.37 ml, 143.5 μl, 1.8 mmol, 4 equivs) inTHF (2 ml). After 4 hours, the solvent was removed under reducedpressure. The gum was dissolved in chloroform (10 ml), and washed with1M HCl (8 ml), sodium bicarbonate (10 ml) and water (15 ml). The organicphase was dried, and the solvent removed in vacuo. The residue waspurified by column chromatography on silica with elution bychloroform-methanol (97:3). Pooling and evaporation of the eluent gavethe product as a white solid.

Spectral Data

323—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(p-ethylphenyl methoxyalaninyl) Phosphoramidate

Yield=79% ³¹P (CDCl₃): 3.43 ppm ¹H (CDCl₃): 9.25 (0.5H, s, B, NH), 9.23(0.5H, S, A, NH), 7.34 (0.5H, s, H-6, B), 7.33 (0.5H, s, H-6, A),7.14–7.00 (5H, m, Ph, H-1′), 6.28 (1H, m, H-3′), 5.88 (1H, m, H-2′),5.00 (1H, m, H-4′), 4.38–4.25 (2H, m, H-5′), 3.93 (2H, m, ala-NH,ala-CH), 3.70 (1.5H, s, OMe, A), 3.67 (1.5H, S, OMe, B), 2.60 (2H, q,CH₂CH₃, J=7.5 Hz), 1.84 (1.5H, d, 5-CH₃, J=1.2 Hz), 1.80 (1.5H, d,5-CH₃, J=1.2 Hz), 1.31 (3H, m, CH₂CH₃), 1.19 (3H, m, ala-CH₃). ¹³C(CDCl₃): 174.25 (ala-CO, A), 174.12 (ala-CO, B), 164.22 (C-4, B), 164.17(C-4, A), 151.15 (C-2, B), 151.12 (C-2, A), 148.29 (i-Ph, B), 148.16(i-Ph, A), 141.24 (p-Ph, A), 141.19 (p-Ph, B), 136.06 (C-6, B), 135.76(C-6, A), 133.50 (C-3′, A), 133.15 (C-3′, B), 129.11 (o-Ph, A), 129.05(o-Ph, B), 127.54 (C-2′, A), 127.36 (C-2′, B), 120.08 (d, m-Ph, B, J=3.9Hz), 119.90 (d, m-Ph, A, J=4.9 Hz), 111.51 (C-5, A), 111.40 (C-5, B),89.83 (C-1′, B), 89.60 (C-1′, A), 84.88 (d, C-4′, B, J=8.8 Hz), 84.70(d, C-4′, A, J=8.8 Hz), 67.11 (d, C-5′, A, J=4.9 Hz), 66.48 (d, C-5′, B,J=4.9 Hz), 52.65 (OMe), 50.26 (ala-CH, B), 50.13 (ala-CH, A), 28.19(Ph-CH₂), 20.97 (d, ala-CH₃, B, J=4.9 Hz), 20.90 (d, ala-CH₃, A, J=4.9Hz), 15.69 (Ph-CH₂CH₃), 12.45 (5-CH₃, A), 12.41 (5-CH₃, B). MS:C₂₂H₂₉N₃O₈P: 494 (MH⁺, 5), 368 (MH⁺-thymine, 25), 228 (15), 81 (C₅H₅O,base peak) Accurate mass: expected 494.1692; found 494.1693 HPLC:RT=27.23 and 27.48 min

324—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenyl N-methoxyalaninyl) phosphoramidate

Yield=88% ³¹P (CDCl₃): 3.20 and 3.86 ppm ¹H (CDCl₃): 1.32 and 1.34 (d,3H, J=6.8 Hz, CH₃ ala); 1.81 and 1.84 (d, 3H, 5CH₃); 3.69 and 3.70 (s,3H, OMe); 3.84–4.00 (m, 2H, CH ala+NH ala); 4.32 (m, 2H, H5′); 5.02 (m,1H, H4′); 5.88 (m, 1H, H2′); 6.33 (m, 1H, H3′); 7.03 (m, 1H, H1′);7.15–7.35 (m, 6H, Ar+H6); 9.22 and 9.26 (bs, 1H, NH) ¹³C (CDCl₃): 12.52(5CH₃); 21.02 (CH₃ ala); 50.22–50.35 (CH ala); 52.74 (OMe); 66.62–67.29(C5′); 84.80–84.88 (C4′); 89.69–89.93 (C1′); 111.44–111.57 (C5);120.13–120.31 (Ar ortho); 125.30 (Ar para); 127.49–127.65 (C2′);129.87–129.93 (Ar meta); 133.19–133.50 (C3′); 135.77–136.06 (C6); 150.51(Ar ipso); 151.16 (C2); 164.14 (C4); 174.12 (CO ala) MS: 466 (MH⁺°, 7);340 (MH⁺°-base); 200 (17); 136 (47); 89 (25); 81 (C₅H₅O, base peak)HPLC: RT=22.48 and 22.87 min

327—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(p-fluorophenyl methoxyalaninyl) phosphoramidate

Yield=89% ³¹P (CDCl₃): 3.16 ppm ¹H (CDCl₃): 9.75 (1H, s, NH), 7.24(0.5H, d, H-6, B, J=1.2 Hz), 7.17 (0.5H, d, H-6, A, J=1.2 Hz), 7.09 (5H,m, Ph, H-1′), 6.22 (1H, m, H-3′), 5.82 (1H, m, H-2′), 4.94 (1H, m,H-4′), 4.30–3.84 (4H, m, ala-NH, ala-CH, H-5′), 3.63 (1.5H, s, OMe, A),3.62 (1.5H, s, OMe, B), 1.77 (1.5H, d, 5-CH₃, B, J=1.0 Hz), 1.74 (1.5H,d, 5-CH₃, A, J=1.0 Hz), 1.29 (1.5H, d, ala-CH₃, B, J=7.0 Hz), 1.23(1.5H, d, ala-CH₃, A, J=7.0 Hz). ¹³C (CDCl₃): 174.19 (d, ala-CO, B,J=6.8 Hz), 174.00 (d, ala-CO, A, J=6.8 Hz), 164.25 (C-4, B), 164.20(C-4, A), 159.77 (d, p-Ph, J=243.6 Hz), 151.14 (C-2), 146.25 (i-Ph),135.99 (C-6, A), 135.70 (C-6, B), 133.40.(C-3′, A), 133.05 (C-3′, B),127.61 (C-2′, B), 127.45 (C-2′, A), 121.70 (m, o-Ph), 116.37 (d, m-Ph,A, J-23.5 Hz), 116.34 (d, m-Ph, B, J=23.5 Hz), 111.45 (C-5, A), 111.32(C-5, B), 89.87 (C-1′, A), 89.63 (C-1′, B), 84.66 (d, C-4′, J=5.9 Hz),67.29 (d, C-5′, A, J=4.9 Hz), 66.10 (d, C-5′, B, J=4.9 Hz), 52.70 (OMe),50.26 (ala-CH, A), 50.13 (ala-CH, B), 20.92 (d, ala-CH₃, A, J=4.8 Hz),20.88 (d, ala-CH₃, B, J=4.8 Hz), 12.45 (5-CH₃, B), 12.41 (5-CH₃, A). MS:C₂₀H₂₄N₃O₈PF : 484 (MH+, 11), 358 (MH⁺-thymine, 20), 218 (13), 154 (32),136 (28), 81 (C₅H₅O, base peak). Accurate mass: expected 484.1285; found484.1318 HPLC: RT=25.17 and 25.40 min

526—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(m-trifluromethylphenylmethoxy alaninyl) phosphoramidate

Yield=80% ³¹P (CDCl₃): 2.49 and 3.16 ppm ¹H (CDCl₃): 9.06 (1H, s, NH),7.45 (5H, m, H-6, Ph), 7.03 (1H, m, H-1′), 6.31 (1H, m, H-3′), 5.92 (1H,m, H-2′), 5.03 (1H, m, H-4′), 4.32 (2H, m, H-5′), 3.97 (2H, m, ala-NH,ala-CH), 3.71 (1.5H, s, OMe, B), 3.70 (1.5H, s, OMe, A), 1.86 (1.5H, s,5-CH₃, B), 1.80 (1.5H, d, 5-CH₃, A), 1.36 (3H, m, ala-CH₃). 13 _(c)(CDCl₃): 174.06 (d, ala-CO, A, J=6.8 Hz), 173.89 (d, ala-CO, B, J=6.8Hz), 163.91 (C-4, A), 163.86 (C-4, B), 150.96 (C-2), 150.71 (d, a-Ph,J=5.9 Hz), 135.86 (C-6, A), 135.66 (C-6, B), 133.30 (C-3′, A), 133.02(C-3′, B), 132.00 (q, c-Ph, J=32.0 Hz), 130.66 (e-Ph), 127.84 (C-2′, B),127.74 (C-2′, A), 123.98 (f-Ph, A), 123.84 (q, CF₃, J=272.0 Hz), 123.79(f-Ph, B), 122.14 (d-Ph), 117.54 (d, b-Ph, J=3.9 Hz), 111.61 (C-5, B),111.44 (C-5, A), 90.04 (C-1′, B), 89.77 (C-1′, A), 84.61 (d, C-4′, J=7.8Hz), 67.60 (d, C-5′, B, J=4.9 Hz), 66.89 (d, C-5′, A, J=4.9 Hz), 52.87(OMe), 50.32 (d, ala-CH, A, J=4.8 Hz), 50.26 (d, ala-CH, B, J=4.8 Hz),21.11 (d, ala-CH₃, B, J=4.9 Hz), 20.99 (d, ala-CH₃, A, J=4.9 Hz), 12.55(5-CH₃, B), 12.47 (5-CH₃, A). MS: C₂₁H₂₄N₃O₈PF₃: 534 (MH⁺, 6), 408(MH⁺-thymine, 8), 268 (10), 149 (10), 81 (C₅H₅O, base peak). Accuratemass: expected 534.1253; found 534.1201 HPLC: RT=30.56 min

546—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(3,5-dichlorophenylmethoxy alaninyl) phosphoramidate

Yield=70% ³¹P (CDCl₃): 2.83 and 3.42 ppm ¹H (CDCl₃): 9.74 (1H, s, NH),7.40 (1H, s, H-6), 7.29 (3H, m, Ph), 7.14 (1H, m, H-1′), 6.44 (1H, m,H-3′), 6.04 (1H, m, H-2′), 5.14 (1H, m, H-4′), 4.48–4.07 (5H, m, ala-NH,ala-CH, H-5′), 3.84 (3H, s, OMe), 1.97 (1.5H, s, 5-CH₃, A), 1.92 (1.5H,s, 5-CH₃, B), 1.48 (3H, m, ala-CH₃). ¹³C (CDCl₃): 173.93 (ala-CO),164.09 (C-4), 151.27 (i-Ph), 151.06 (C-2), 136.01 (m-Ph), 135.60 (C-6),133.14 (C-3′, B), 132.89 (C-3′, A), 127.83 (C-2′), 125.69 (p-Ph), 119.40(o-Ph), 111.54 (C-5, A), 111.40 (C-5, B), 90.03 (C-1′, A), 89.74 (C-1′,B), 84.60 (C-4′), 67.68 (C-5′, A), 66.98 (C-5′, B), 52.85 (OMe), 50.26(ala-CH), 20.93 (ala-CH₃), 12.51 (5-CH₃). MS: C₂₀H₂₃N₃O₈PCl₂: 534 (MH⁺,8), 408 (MH⁺-thymine, 12), 391 (10), 149 (12), 127 (thymineH+, 12), 81(C₅H₅O, base peak). Accurate mass: expected 534.0600; found 534.0589HPLC: RT=32.19 min

730—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenyl N-benzyloxyalaninyl) phosphoramidate

Yield=92% ³¹P (CDCl₃): 3.40 and 4.04 ppm ¹H (CDCl₃): 1.24 and 1.26 (d,3H, J=6.8 Hz, CH₃ ala); 1.70 and 1.74 (s, 3H, 5CH₃); 3.86–4.28 (m, 4H,H5′+CH ala+NH); 4. 85 (m, 1H, H4′); 5.04 and 5.06 (s, 2H, CH₂Ph); 5.74(d, 1H, H2′); 6.16 (dd, 1H, H3′); 6.90 (m, 1H, H1′); 7.00–7.30 (m, 11H,Ar+H6); 9.61 (d, 1H, NH) ¹³C (CDCl₃): 12.52 (5CH₃); 20.98 (CH₃ ala);50.36–50.52 (CH ala); 66.70–67.18 (C5′); 67.46 (CH₂Ph);84.63–84.76–84.88 (C4′); 89.68–89.88 (C1′); 111.44–111.55 (C5);120.18–120.25–120.36–120.43 (Ar ortho, OPh); 125.31 (Ar para, OPh);127.48–127.61 (C2′); 128.45–128.79–128.83 (Ar, CH₂Ph); 129.87–129.93 (Armeta, OPh); 133.16–133.45 (C3′); 135.35 (Ar1, CH₂Ph); 135.79–136.07(C6); 150.44 (Ar1, OPh); 151.18 (C2); 164.21–164.28 (C4);173.42–173.51–173.65 (CO ala) HPLC: RT=34.96 and 35.07 min MS:C₂₆H₂₈O₈N₃P: 542(MH⁺°; 17); 416 (MH⁺°base; 40); 81(100). Accurate mass:expected 542.1716; found 542.1712

776—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(2,4-dibromophenylN-methylalaninyl) phosphoramidate

Yield=88% ³¹P (CDCl₃): 3.07 and 3.62 ppm ¹H (CDCl₃): 1.26 and 1.28 (d,3H, J=6.8 Hz, CH₃ ala); 1.75 and 1.80 (s, 3H, 5CH₃); 2.11 (s, 1H, NH);3.64 (s, 3H, OMe); 3.92–4.30 (m, 3H, H5′+CHala); 4.98 (m, 1H, H4′); 5.87(m, 1H, H2′); 6.26 (m, 1H, H3′); 6.96 (m, 1H, H1′); 7.30–7.60 (m, 4H,Ar+H6); 9.41(bs, 1H, NH) ¹³C (CDCl₃): 12.51 (5CH₃); 21.00 (CH₃ ala);50.24 (CHala); 52.80 (OMe); 67.37–67.83 (C5′); 84.49–84.61 (C4′);89.80–89.92 (C1′); 111.60 (C5); 115.49 (Ar2); 118.26 (Ar4);122.61–122.89 (Ar6); 127.70 (C2′); 131.86 (Ar5); 133.06–133.21 (C3′);135.64 (Ar3); 135.75–135.88 (C6); 147.01 (Ar1); 151.07 (C2); 164.03(C4); 173.71–173.82 (COala) HPLC: RT=41.17 and 41.30 min MS:C₂₀H₂₂O₈N₃PBr₂: 622,624,626 (MH⁺°; 3,6,3); 496,498,500 (MH⁺°^(n−)base;5,9,5); 81 (100). Accurate mass: expected 621.9516; found 621.9507

779—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(2,3,4,5,6-pentafluorophenyl-N-methylalaninyl)phosphoramidate

Yield=76% ³¹P (CDCl₃): 4.74 and 5.66 ppm ¹H (CDCl₃): 1.34 and 1.36 (d,3H, J=6.7 Hz, CH₃ ala); 1.75 and 1.81 (s, 3H, 5CH₃); 3.69 (s, 3H, OMe);3.92–4.40 (m, 4H, H5′+CH ala+NH); 4.97 (m, 1H, H4′); 5.85 (m, 1H, H2′);6.29 (m, 1H, H3′); 6.93 (m, 1H, H1′); 7.19 (m, 1H, H6); 9.38 (bs, 1H,NH) ¹³C (CDCl₃): 12.23–12.43 (5CH₃); 20.83 (CH₃ ala); 50.22–50.34 (CHala); 52.99 (OMe); 67.75–68.37 (C5′); 84.42–84.52 (C4′); 89.87–90.17(C1′); 111.75 (C5); 127.69–127.93 (C2′); 132.86–133.13 (C3′); 132–143(m, Ar); 135.74–135.96 (C6); 151.11 (C2); 164.15 (C4); 173.64–173.76(COala) Mass (NOBA matrix): C₂₀H₁₉O₈N₃PF₅: 556 (MH⁺°, 31); 578 (M°⁺Na,100) HPLC: RT=35.90 min

862—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenyl N-hexyloxyalaninyl) phosphoramidate

Yield=88% ³¹P (CDCl₃): 3.99 and 4.60 ppm ¹H (CDCl₃): 0.94 (m, 3H, CH₃CH₂); 1.28–1.41 (m, 9H, CH₃ ala+3×CH₂); 1.65 (m, 2H, CO₂CH₂CH ₂); 1.90and 1.93 (s, 3H, 5CH₃); 4.00–4.20 (m, 4H, CH ala+NH ala+CO₂CH ₂); 4.37(m, 2H, H5′); 5.05 (m, 1H, H4′); 5.94 (m, 1H, H2′); 6.38 (m, 1H, H3′);7.10 (m, 1H, H1′); 7.15–7.36 (m, 6H, Ar+H6); 9.48 and 9.51 (s, 1H, NH)¹³C (CDCl₃): 12.76 (5CH₃); 14.39 (CH₃CH₂); 21.45 (CH₃ ala); 22.88,25.82, 28.82 and 31.72 (CH₂); 50.63 (CH ala); 66.26 (OCH₂); 66.89–67.43(C5′); 85.03 (C4′); 89.97 (C1′); 111.68–111.83 (C5); 120.55 (Ar ortho);125.57 (Ar para); 127.86 (C2′); 130.15 (Ar meta); 133.47–133.70 (C3′);136.03–136.31 (C6); 150.72 (Ar ipso); 151.37–151.39 (C2); 164.35–164.42(C4); 174.02 (CO ala) Mass (NOBA matrix): C₂₅H₃₄O₈N₃P: 536 (MH⁺°, 24);558 (M°⁺Na, 37)

863—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenylN-methoxy-phenylalaninyl) phosphoramidate

Yield=89% ³¹P (CDCl₃): 3.96 and 4.35 ppm ¹H (CDCl₃): 1.89 (s, 3H, 5CH₃);3.00 (m, 2H, CH₂Ph); 3.74 (s, 3H, OMe); 3.80–4.28 (m, 4H, CH ala+NHala+H5′); 4.94 (m, 1H, H4′); 5.91 (m, 1H, H2′); 6.21–6.30 (m, 1H, H3′);7.04–7.32 (m, 12H, Ar+H1′+H6); 9.35 (s, 1H, NH) ¹³C (CDCl₃): 12.54(5CH₃); 40.55 (CH₂Ph); 52.63 (OMe); 55.72–56.01 (CH ala); 66.50–67.10(C5′); 84.78 (C4′); 89.71–89.95 (C1′); 111.53–111.64 (C5); 120.28 (Arortho, OPh); 125.40 (Ar para, OPh); 127.52 (C2′); 128.86, 129.65 and129.98 (Ar, CH₂Ph); 129.86–129.92 (Ar meta, OPh); 133.18–133.50 (C3′);135.72 (Ar ipso, CH₂Ph); 135.79–136.06 (C6); 150.46 (Ar ipso, OPh);151.13–151.17 (C2); 164.12–164.18 (C4); 173.00 (CO ala) Mass (NOBAmatrix): C₂₆H₂₈O₈N₃P: 542 (MH³⁰ °,77); 564 (M°⁺Na, 29)

864—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenylN-methoxy-leucinyl) phosphoramidate

Yield=87% ³¹P (CDCl₃): 4.18 and 4.83 ppm ¹H (CDCl₃): 0.91 (m, 6H, (CH₃)₂CH); 1.42–1.70 (m, 3H, CH ₂CH(CH₃)₂); 1.91 and 1.93 (s, 3H, 5CH₃);3.73 (s, 3H, OMe); 3.76–3.98 (m, 2H, CH ala+NH ala); 4.28–4.46 (m, 2H,H5′); 5.08 (m, 1H, H4′); 5.96 (m, 1H, H2′); 6.36 (m, 1H, H3′); 7.09 (m,1H, H1′); 7.18–7.35 (m, 6H, Ar+H6); 9.35 (s, 1H, NH) ¹³C (CDCl₃): 12.76(5CH₃); 22.23–23.01 ((CH₃)₂CH); 24.75 (CH(CH₃)₂); 43.86–44.11(CH₂CH(CH₃)₂); 52.75 (OMe); 53.42–53.60 (CH ala); 66.92–67.55 (C5′);85.62 (C4′); 89.92–90.19 (C1′); 111.69–111.83 (C5); 120.37–120.62 (Arortho); 125.55–125.58 (Ar para); 127.79 (C2′); 130.12 (Ar meta);133.51–133.70 (C3′); 136.00–136.36 (C6); 151.05 (Ar ipso); 151.38 (C2);164.39–164.50 (C4); 174.55–174.88 (CO ala) Mass (NOBA matrix):C₂₃H₃₀O₈N₃P: 508 (MH⁺°, 62); 530 (M°⁺Na, 59)

865—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenyl N-methoxyvalinyl)phosphoramidate

Yield=86% ³¹P (CDCl₃): 4.85 and 5.40 ppm ¹H (CDCl₃): 0.92 (m, 6H, (CH₃)₂CH); 1.82 (m, 3H, CH(CH₃) ₂); 1.89 and 1.91 (s, 3H, 5CH₃); 3.76 (s,3H, OMe); 3.82 (m, 2H, CH ala+NH ala); 4.30–4.48 (m, 2H, H5′); 5.07 (m,1H, H4′); 5.96 (m, 1H, H2′); 6.38 (m, 1H, H3′); 7.10 (m, 1H, H1′);7.18–7.35 (m, 6H, Ar+H6); 9.31 (s, 1H, NH) ¹³C (CDCl₃): 12.80 (5CH₃);17.77–19.24 ((CH₃)₂CH); 32.43–32.62(CH(CH₃)₂); 52.67 (OMe); 60.32–60.38(CH ala); 66.92–67.65 (C5′); 85.04 (C4′); 89.98–90.24 (C1′);111.76–111.87 (C5); 120.45–120.56 (Ar ortho); 125.54–125.59 (Ar para);127.81–127.86 (C2′); 130.13–130.17 (Ar meta); 133.51–133.72 (C3′);136.01–136.28 (C6); 150.83 (Ar ipso); 150.87–151.34 (C2); 164.30–164.37(C4); 173.56–173.65 (CO ala) Mass: C₂₂H₂₈O₈N₃P: 493.6 (MH⁺°, 100)

866—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenyl N-methoxyglycinyl)phosphoramidate

Yield=90% ³¹P (CDCl₃): 4.89 and 5.52 ppm ¹H (CDCl₃): 1.79 and 1.83 (s,3H, 5CH₃); 3.69 (s, 3H, OMe); 3.70–4.05 (m, 4H, CH₂NH+CH ala+NH ala);4.32 (m, 2H, H5′); 4.99 (m, 1H, H4′); 5.92 (m, 1H, H2′); 6.38 (m, 1H,H3′); 6.98 (m, 1H, H1′); 7.05–7.38 (m, 6H, Ar+H6); 9.44 and 9.46 (s, 1H,NH) ¹³C (CDCl₃): 12.75 (5CH₃); 43.15 (CH₂NH); 52.94 (OMe); 66.78–67.52(C5′); 84.98–85.10 (C4′); 89.68–90.16 (C1′); 111.69–111.80 (C5);120.46–120.59 (Ar ortho); 125.66 (Ar para); 127.66–127.91 (C2′); 130.22(Ar meta); 133.48–133.87 (C3′); 136.11–136.40 (C6); 150.65 (Ar ipso);151.45 (C2); 164.46 (C4); 171.41–171.51 (CO ala) Mass (NOBA matrix):C₁₉H₂₂O₈N₃P: 452 (MH⁺°, 74); 474 (M°⁺Na, 46)

867—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenylN-methoxymethioninyl) phosphoramidate

Yield=81% ³¹P (CDCl₃): 4.09 and 4.86 ppm ¹H (CDCl₃): 1.74 and 1.79 (s,3H, CH₃S); 1.94 and 1.97 (s, 3H, 5CH₃); 1.80–2.40 (m, 5H, CHCH ₂CH ₂S);3.72 and 3.74 (s, 3H, OMe); 3.98–4.32 (m, 4H, H5′+CH ala+NH ala); 4.96(m, 1H, H4′); 5.84 (m, 1H, H2′); 6.26 (m, 1H, H3′); 6.96 (m, 1H, H1′);7.05–7.25 (m, 6H, Ar+H6); 9.58 (bs, 1H, NH) ¹³C (CDCl₃): 12.80 (5CH₃);15.68 (CH₃S); 29.95 (CH₂SCH₃); 33.73–33.85 (CH₂CH₂S);53.06 (OMe);53.81–54.07 (NHCH); 67.05–67.70 (C5′); 84.90–85.03 (C4′); 89.98–90.23(C1′); 111.66–111.86 (C5); 120.39–120.66 (Ar ortho); 125.63 (Ar para);127.81–127.91 (C2′); 130.13 (Ar meta); 133.44–133.69 (C3′);136.00–136.38 (C6); 150.72–150.80 (Ar ipso); 151.41 (C2); 164.52 (C4);173.61–173.94 (CO ala) Mass (NOBA matrix): C₂₂H₂₈O₈N₃PS: 526 (MH⁺°, 46);548 (M°⁺6Na, 21)

868—2′,3′-dideoxy-2′, 3′-didehydrothymidine-5′-(2,4-dibromophenylN-benzylalaninyl) phosphoramidate

Yield=82% ³¹P (CDCl₃): 3.68 and 4.18 ppm ¹H (CDCl₃): 1.40 and 1.42 (d,3H, J=6.7 Hz, CH₃ ala); 1.90 and 1.92 (s, 3H, 5CH₃); 4.04–4.40 (m, 4H,H5′+CHala+NH ala); 4.98 (m, 1H, H4′); 5.20 (s, 2H, CH₂Ph); 5.91 (m, 1H,H2′); 6.27 and 6.35 (m, 1H, H3′); 7.06 (bs, 1H, H1′); 7.30–7.70 (m, 9H,Ar+H6); 9.52 (s, 1H, NH) ¹³C (CDCl₃): 12.86 (5CH₃); 21.35 (CH₃ ala);50.68–50.76 (CHala); 67.67–68.03 (C5′); 67.88 (CH₂Ph); 84.85 (C4′);90.10–90.20 (C1′); 111.88–111.92 (C5); 115.76–115.91 (Ar2);118.62–118.72 (Ar4); 122.91–123.22 (Ar6); 127.98 (C2′);128.75–129.01–129.12 (Ar o,m,p, CH₂Ph); 132.20 (Ar5); 133.38–133.51(C3′); 135.48 (Ar ipso, CH₂Ph); 135.96 (Ar3); 136.21 (C6); 147.28 (Ar1);151.39 (C2); 164.34–164.38 (C4); 173.47–173.62 (COala) Mass (NOBAmatrix): C₂₆H₂₆O₈N₃PBr₂: 699–700–701 (MH⁺°, 27-49-29); 721-722-723(M°⁺Na, 17-21-17)

877—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenyl N-methoxyglycinyl)phosphoramidate

Yield=83% ³¹P (CDCl₃): 3.91 and 4.33 ppm ¹H (CDCl₃): 1.83 and 1.85 (s,3H, 5CH₃); 3.01 (m, 2H, CHCH ₂Ph); 3.78–4.30 (m, 4H, H5′+HNCH); 4.92 (m,1H, H4′); 5.89 (m, 1H, H2′); 6.18 and 6.27 (m, 1H, H3′); 7.00–7.40 (m,17H, Ar+H1′+H6); 9.35 (bs, 1H, NH) ¹³C (CDCl₃): 12.62–12.75 (5CH₃);40.65–40.73 (CHCH₂Ph); 55.95–56.26 (NHCH); 66.79–67.27 (C5′); 67.80(CH₂Ph); 84.87–85.05 (C4′); 89.92–90.14 (C1′); 111.72–111.82 (C5);120.45–120.52 (Ar ortho, OPh); 125.60 (Ar para, OPh); 127.73 (C2′);129.01–129.07–129.11–129.91–130.15–130.38–135.29–135.85 (Ar, 2×CH₂Ph);130.21 (Ar meta, OPh); 133.36–133.63 (C3′); 136.24 (C6); 150.68–150.77(Ar ipso, OPh); 151.31–151.35 (C2); 164.28–164.34 (C4); 172.48–172.64(CO ala) Mass (NOBA matrix): C₃₂H₃₂O₈N₃P: 618 (MH⁺°, 78); 640 (M°⁺Na,52)

878—2′,3′-dideoxy-2′,3′-didehydrothymidine-5′-(phenylN-tert-butylphenylalaninyl) phosphoramidate

Yield=79% ³¹P (CDCl₃): 4.27 and 4.50 ppm ¹H (CDCl₃): 1.40 and 1.41 (s,9H, tBu); 1.84 and 1.87 (s, 3H, 5CH₃); 3.00 (m, 2H, CH ₂Ph); 3.76–4.28(m, 4H, H5′+HNCH); 4.95 (m, 1H, H4′); 5.86 and 5.91 (m, 1H, H2′); 6.26and 6.30 (m, 1H, H3′); 7.04 (m, 1H, H1′); 7.12–7.25 (m, 11H, Ar+H6);9.38 and 9.40 (bs, 1H, NH) ¹³C (CDCl₃): 12.76–12.79 (5CH₃); 28.31((CH₃)₃C); 40.96–41.04 (CH₂Ph); 56.31–56.65 (NHCH); 66.79–67.28 (C5′);82.90–82.92 ((CH₃)₃ C); 84.94–85.03 (C4′); 89.93–90.11 (C1′);111.67–111.86 (C5); 120.45 (Ar ortho, OPh); 125.52 (Ar para, OPh);127.77 (C2′); 127.88–128.83–128.92–136.02 (Ar, CH₂ Ph); 130.13 (Ar meta,OPh); 133.54–133.60 (C3′); 136.31 (C6); 150.75–150.84 (Ar ipso, OPh);151.36 (C2); 164.32–164.37 (C4); 171.89 (CO ala) Mass (NOBA matrix):C₂₉H₃₄O₈N₃P: 584 (MH⁺°, 26); 606 (M°⁺Na, 41)

892—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenyl N-cyclohexyloxyalaninyl) phosphoramidate

Yield=83% ³¹P (CDCl₃): 4.11 and 4.71 ppm. ¹H (CDCl₃): 1.08–1.82 (m, 16H,CH₃ ala+5CH₃+cyclohexyl); 3.79–4.14 (m, 2H, CH ala+NH ala); 4.27 (m, 2H,H5′); 4.69 (m, CH cyclohexyl); 4.96 (m, 1H, H4′); 5.80 (m, 1H, H2′);6.24 (m, 1H, H3′); 6.98 (m, 1H, H1′); 7.04–7.32 (m, 6H, Ar+H6); 9.66 and9.82 (bs, 1H, NH). ¹³C (CDCl₃): 12.58 (5CH₃); 21.18–21.32 (CH₃ ala);23.73–25.40–31.49–31.58(CH₂ cyclohexyl); 50.47–50.61 (CH ala);66.69–67.24 (C5′); 74.36(CH cyclohexyl); 84.87 (C4′); 89.72–89.92 (C1′);111.48–111.63 (C5); 120.26–120.49 (Ar ortho); 125.32–125.37 (Ar para);127.59–127.73 (C2′); 129.91–129.98 (Ar meta); 133.30–133.51 (C3′);135.89–136.16 (C6); 150.53 (Ar ipso); 150.67–151.31(C2) 164.36–164.41(C4); 173.23 (CO ala). Mass (NOBA matrix): C₂₅H₃₂O₈N₃P: 534 (MH⁺°, 56);556 (M°⁺Na, 42)

893—2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenyl N-tButyloxyalaninyl) phosphoramidate

Yield=79% ³¹P (CDCl₃): 4.17 and 4.67 ppm. ¹H (CDCl₃): 1.34 (m, 3H, CH₃ala); 1.46 (m, 9H, CH₃ tBu); 1.87 (d, 3H, 5CH₃); 3.82–4.06 (m, 2H, H5′);4.29–4.49 (m, 2H, CH ala+NH ala); 5.05 (m, 1H, H4′); 5.91 (m, 1H, H2′);6.35 (m, 1H, H3′); 7.06 (m, 1H, H1′); 7.15–7.40 (m, 6H, Ar+H6); 9.60(bs, 1H, NH).

¹³C (CDCl₃): 12.54 (5CH₃); 21.19–21.35 (CH₃ ala); 28.07 (C(CH₃)₃);50.80–50.89 (CH ala); 66.60–67.18 (C5′); 82.41–82.45(C(Me)₃); 84.82(C4′); 89.67–89.87 (C1′); 111.44–111.60 (C5); 120.22–120.41 (Ar ortho);125.28–125.31 (Ar para); 127.54–127.65 (C2′); 129.88–129.94 (Ar meta);133.33–133.47 (C3′); 135.84–136.10 (C6); 150.51 (Ar ipso); 150.65–151.20(C4); 164.19–164.23 (C2); 172.78–172.93 (CO ala). Mass (NOBA matrix):C₂₃H₃₀O₈N₃P: 508 (MH⁺°, 82); 530 (M°⁺Na, 48).

2′,3′-Dideoxy-2′,3′-didhydrothymidine-5′-(phenyl methoxy-B-alaninyl)phosphate

Cf 1197

Yield=64% ³¹P (CDCl₃): 6.44, 6.70(1:3) 1H (CDCl₃): 1.87° (s, 3H, 5-CH₃),2.42(t, 2H, CH₂ ala), 3.22° (m, 2H, CH₂ ala), 3.62 (s, 3H, OCH₃), 4.09(m, 1H, H4′), 4.18–4.39 (m, 2H, H5′), 4.97 (bs, 1H, NH ala), 5.88° (m,1H, H2′), 6.32° (m, 1H, H3′), 6.99 (m, 1H, H1′), 7.08–7.38 (m, 5H, Phand H6), 10.01 (bs, 1H, base NH) ¹³C (CDCl₃): 14.52 (5-CH₃), 37.80° (CH₂ala), 39.28° (CH₂ ala) 53.91° (OCH₃), 68.57° (d, J=3.92 Hz, C5′), 86.90(d, J=8.38 Hz, C4′), 91.68° (C1′), 113.40° (C5), 122.34 (d, J=4.68 Hz,ortho-Ph), 127.23 (C2′), 129.55° (para-Ph), 131.81^(°)(meta-Ph), 135.45°(C6), 137.99° (C3′), 152.60° (d, J=5.96 Hz, ipso-Ph), 153.44 (C2),166.58 (C4), 174.55° (COO) Mass (NOBA matrix): C₂₀H₂₄N₃O₈P 126(thymine⁺,5), 127 (thymineH⁺,4), 242 (C₁₀H₁₃PO₄N⁺,9) 243(C₁₀H₁₄PO₄N⁺,3), 465 (M⁺,4), 466 (MH⁺, 8), 467 (MHNa⁺, 20), 168 (MHNa⁺,¹³O, 5), 187 (MNa⁺, 3), 188 (MHNa⁺, 97), 189 (MHNa⁺, ¹³C, 21) HighResolution MS: found 466.1379 (MH⁺), C₂₀H₂₅N₃O₈P requires 466.1379 HPLC:RT=22.81, 23.27 mins (1:1)

2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(phenylmethoxy-α-aminobutylryl)phosphate

Cf 1198 Yield=65% ³¹P (CDCl₃): 6.11, 6.66 (1:2) ¹H (CDCl₃): 1.78 (m, 2H,CH₂ GABA), 1.85° (s, 3H, 5-CH₃), 2.35 (t, 2H, J=6.95 Hz, CH₂ GABA),2.97° (m, 2H, CH₂ GABA), 3.68 (s, 3H, OCH₃), 3.93° (m, 1H, H4′), 4.28°(m, 1H, H5′), 4.35^(°)(m, 1H, H5′), 5.02 (bs, 1H, NH GABA), 5.82° (m,1H, H2′), 6.31 (m, 1H, H3′), 6.98 (m, 1H, H1′), 7.11–7.37 (m, 6H, Ph andH6), 9.91 (bs, 1H, base NH) ¹³C(CDCl₃): 12.64 (5-CH₃), 26.72° (CH₂GABA), 32.25° (CH₂ GAEA) 40.98° (CH₂ GABA), 51.94 (OCH₃), 66.93° (C5′),85.11 (d, J=8.30 Hz, C4′), 111.40 (C5), 120.46° (d, J=4.83 Hz,ortho-PH), 125.24 (C2′), 127.59° (para-Ph), 129.88° (meta-Ph), 133.68°(C6), 136.28° (C3′), 150.86° (d, J=6.45 Hz, ipso-Ph), 151.61 (C2),164.80 (C4), 173.86 (COO) Mass (matrix NOBA): C₂₁H₂₆N₃O₈P: 127(thymineH⁺, 28), 479 (M⁺, 3), 480 (MH⁺, 59), 481 (MH⁺, ¹³C, 17), 501(MNa⁺, 3), 502 (MHNa⁺, 59), 503 (MHNa⁺, ¹³C, 16) High Resolution MS:found 480.1486 (MH⁺), C₂₁H₂₇N₃O₈P requires 480.1536 HPLC: RT 23.90,24.33 mins (1:1)

2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(phenylmethoxy-2-aminoisobutylryl) phosphate

Cf 1200

Yield=36% ³¹P (CDCl₃): 2.38, 3.05 (3:1) ¹H (CDCl₃): 1.53° (s, 6H, CMe₂),1.91° (s, 3H, 5-CH₃), 3.71 (s 3H, OCH₃), 4.31 (m, 2H, H5′), 4.23–4.41(m, 3H, H4′ and H5′), 5.03 (bs, 1H, P—NH) 5.89° (m, 1H, H2′), 6.28° (m,1H, 3′), 6.99–7.31 (m, 7H, Ph, HO and H1′), 9.09 (bs, 1H, base NH) ¹³C(CDCl₃): 14.27 (5-CH₃), 28.74 (CMe ₂), 54.81° (OCH₃), 58. (CMe₂), 69.03°(d, C′, J=5.58 Hz), 86.57° (d, J=7.88 Hz, C4′), 91.51° (C1′), 113.24°(C5), 122.01° (d, J=4.95 Hz, ortho-Ph), 126.88 (C2′), 129.25° (para-Ph),131.57° (meta-Ph), 135.19° (C6), 137.68° (C3′), 152.52° (d, J=3.09 Hz,ortho-Ph), 153.05 (C2), 166.12 (C4), 177.69° (COO) MS (matrix NOBA): 354((MH-thymine)⁺, base peak), 479 (M⁺, 3), 480 (MH⁺, 64), 481 (MH⁺, ¹³C,17), 482 (MH⁺, 2×¹³C, 3), 502 (MNa⁺, 92), 503 (MHNa⁺, 24) HighResolution MS: found 480.1503 (MH⁺), C₂₁H₂₇N₃O₈P requires 480.1536 HPLC:RT 24.79, 25.29 mins (1:1)

2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(phenylmethoxy-6-aminocaproyl) phosphate

Cf 1199

Yield=80% ³¹P (CDCl₃): 6.90, 6.30 (1:1) ¹H (CDCl₃): 1.28 (s, 2H, CH₂caproyl), 1.45 (m, 2H, CH₂ caproyl), 1.58 (m, 2H, CH₂ caproyl), 1.82°(s, 3H, 5-CH₃), 2.28 (m, 2H, CH₂ caproyl), 2.87 (m, 2H, CH₂ caproyl),3.65 (s, 3H, OCH₃), 3.81 (m, 1H, H4′), 4.25 (m, 2H, H5′), 4.95 (bs, 1H,NH caproyl), 5.86° (m, 1H, H2′), 6.31° (m, 1H, H3′), 6.98 (m, 1H H1′),7.04–7.38° (m, 6H, Ph and H6), 10.12 (bs, 1H, base NH) ¹³C (CDCl₃):13.47° (5-CH₃), 25.43° (CH₂ caproyl), 27.04° (CH₂ caproyl), 32.15° (CH₂caproyl), 34.85 (CH₂ caproyl), 42.30° (CH₂ caproyl), 52.61 (OCH₃),67.92° (C5′), 85.80 (d, J=8.22 Hz), 90.68° (C4′), 112.25° (C5), 121.17°(d, J=4.58 Hz, ortho-Ph), 125.99 (C2′), 128.40° (para-Ph), 130.77(meta-Ph), 134.38° (C6), 137.09° (C3′), 151.69° (d, J=3.23 Hz,ortho-Ph), 152.26 (C2′), 165.36 (C4′), 175.07 (COO) MS (matrix Cl): 127(thymineH⁺, 42), 508 (MH⁺, 18), 509 (MH⁺, ¹³C, 5) High Resolution MS:found 508.1850 (MH⁺), C₂₃H₃₁N₃O₈P requires 508.1849 HPLC: RT 26.33 mins

2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(β-alaninyl) phosphateammonium salt

Cf1216

Yield=62% ³¹P (D₂O): 8.84 ¹H (D₂O): 1.73 (3H, s, 5-CH₃), 2.18 (2H, m,ala CH₂), 2.65 (m, 2H, ala CH₂), 3.79 (2m, H, H5′), 4.95 (m, 1H, H4′),5.76 (m, 1H, H2′), 6.35 (m, 1H, H3′), 6.82 (m, 1H, H1′), 7.47 (s, 1H,H6) ¹³C (D₂O): 11.81 (5-CH₃), 38.51 (ala CH₂), 39.45 (d, ala CH₂, J=6.64Hz), 65.41 (d, C5′, J=4.91 Hz), 86.40 (d, J=9.20 Hz, C4′), 90.20 (C1′),111.07 (C5), 125.40 (C2′), 134.66 (C3′), 138.54 (C6), 152.53 (C2),167.00 (C4), 181,04 (COO) HPLC: RT=32.74 mins.

2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′(γ-aminobutylryl) phosphateammonium salt

Cf 1224 Yield=54% ³¹P (D₂O): 10.03 ¹H(D₂O): 1.47 (m, 2H, GABA CH₂), 1.72(s, 3H, 5-CH3), 1.98 (m, 2H, GABA CH₂), 2.48 (m, 2H, GABA CH₂), 3.72 (m,2H, H5′), 4.91 (m, 1H, H4′), 5.72 (m, 1H, H2′), 6.26 (m, 1H, H3′), 6.72(m, 1H, H1′), 7.45 (s, 1H, H6′). ¹³C (D₂O): 11.79 (5-CH₃), 27.99 (d,J=7.25 Hz, GABA CH₂) 34.47 (GABA CH₂), 41.17 (GABA CH₂), 65.35 (d,J=4.68 Hz, C5′), 86.38 (d, J=9.36 Hz, C4′), 90.27 (C1′), 111.47 (C5′),125.29 (C2′), 134.70 (C3′), 138.68 (C6), 152.47 (C2), 166.95 (C4),182.32 (COO)

2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(caproyl) phosphate ammoniumsalt Cf 1217

Yield=49% ³¹P (D₂O): 10.18 ¹H (D₂O): 1.01 (m, 2H, caproyl CH₂), 1.21 (m,2H, caproyl CH₂), 1.32 (m, 2H, caproyl CH₂), 1.78 (s, 3H, 5-CH₃), 2.05(m, 2H, caproyl CH₂), 2.58 (m, 2H, caproyl CH₂), 3.78 (m, 2H, H5′), 4.99(s, 1H, H4′), 6.32 (m, 1H, H3′), 6.82 (m, 1H, H2′), 7.51 (s, 1H, H6) ¹³C(D₂O): 11.84 (5-CH₃), 25.66 (caproyl CH₂), 26.46 (caproyl CH₂), 31.10(d, J=6.82 Hz, caproyl CH₂), 37.06 (caproyl CH₂), 41.47 (caproyl CH₂),65.37 (d, J=4.83 Hz , C5′), 86.45 (d, J=9.74 Hz, C4′), 90.29 (C1′),111.43 (C5), 125.27 (C2′), 134.80 (C3′), 138.89 (C6), 152.48 (C2),166.94 (C4), 183.15 (COO).

2′,3′-dideoxycytidine-5′-(phenyl-N-methoxyalaninyl) phosphoramidate Cf1221

Yield=16.6% ³¹P (CDCl₃): 3.94, 4.00 ¹H (CDCl₃): 1.33, 1.35 (2×d, 3H, CH₃ala); 1.92, 1.96, 2.41 (1H, 2H, 1H, 3×m, H2′, H3′); 3.66 (s, 3H, OMe);3.86–4.35 (m, 5H, H4′, H5′, CH ala, NH ala); 5.63 (2×d, J=7.4 Hz, H6),6.02 (m, 1H, H-1′), 7.12–7.32 (m, 5H, Ar), 7.73 (1H, 2×d, J=7.4 Hz, H5)¹³C (CDCl₃): 20.98 (CH₃ ala); 24.97, 25.11, 32.85 (C2′, C3′); 50.12,50.30 (CH₃ ala); 52.55 (OMe); 67.19, 67.26, 67.50 (C5′); 79.16, 79.27,79.34 (C4′); 87.29, 87.46 (C1′); 93.48 (C5); 119.99, 120.04, 120.10,125.05, 125.10, 129.73, 129.77 (CAr); 141.17 (C6); 150.48, 150.57 (Cipso Ar); 155.68 (C2); 165.44 (C4); 173.84, 173.94 (COala) Mass (ES⁺):C₁₉H₂₅N₄O₇P: 475 (MNa⁺, 100); HPLC: RT=20.53, 21.22 min

2′,3′-dideoxy-2′,3-didehydrothymidine 5′-(phenylmethoxysarcosinylphosphate) Cf 1098

Yield=65% ³¹P (CDCl₃): 6.80, 7.36 ppm ¹H (CDCl₃): 1.72 (s, 3H, 5CH₃);2.64, 2.67 (s, 3H, NCH₃); 3.62 (s, 3H, OCH₃); 3.40–4.10 (m, 2H, CH₂);4.20–4.50 (m, 2H, H5′); 4.97 (bs, 1H, H4′), 5.80–5.90 (m, 1H, H2′);6.30–6.40 (m, 1H, H3′); 6.97 (bs, 1H, H1′); 7.00–7.30 (m, 6H, Ar+H6);9.59 (bs, 1H, NH) ¹³C (CDCl₃): 12.35 (5CH₃); 34.55–34.60–34.65 (NCH₃);50.67-50.78–50.87 (CH₂); 52.10–52.13 (OCH₃); 62.27–66.77–66.82 (C5′);84.71–84.84 (C4′); 89.52–89.82 (C1′); 111.16–111.33 (C5); 120–150 (m,Ar); 127.17–127.40 (C2 ′); 133.25–133.62 (C3′); 135.73–136.11 (C6);150.85–150.90 (C2); 163.84–163.87 (C4); 170.57–170.60–170.84 (COOCH₃)Mass: C₂₀H₂₄O₈N₃P: 488 ((M+Na)+, 100); 466 ((M+H)⁺, 5) HPLC: RT=25.17and 25.59 min

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenylethoxysarcosinylphosphate) Cf 1133

Yield=65% ³¹P (CDCl₃): 0.87, 7.41 ppm ¹H (CDCl₃): 1.18–1.24 (m, 2H,CH₃CH ₂); 1.80 (s, 3H, 5CH₃); 2.68, 2.71 (s, 3H, NCH₃); 3.46–3.65 (m,2H, NCH₂); 3.91–4.45 (m, 2H, H5′); 4.11, 4.13 (s, 3H, CH₂CH ₃); 5.00(bs, 1H, H4′); 5.82–5.88 (m, 1H, H2′); 6.33–6.37 (m, 1H, H3′); 7.00 (bs,1H, H1′); 7.10–7.50 (m, 6H, Ar+H6); 8.75 (bs, 1H, NH) ¹³C (CDCl₃):12.86–12.89 (5CH₃); 14.69 (CH₂ CH₃); 35.06–35.11 (NCH₃);51.35–51.43–51.51 (NCH₂); 61.77 (CH₂CH₈); 66.77–67.27–67.33 (C5′);85.26–85.36 (C4′); 90.01–90.31 (C1′); 111.69–111.86 (C5); 120–151 (m,Ar); 127.73–127.96 (C2′); 133.73–134.10 (C3′); 136.27–136.64 (C6);151.61 (C2); 164.70 (C4); 170.62–170.66–170.85 (COOCH₃) Mass:C₂₁H₂₆O₈N₃P: 502 ((M+Na)⁺, 100); 480 ((M+H)⁺, 5) HPLC: RT=25.84 and26.65 min

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(methioninyl phosphate) Cf1156

Yield=52% ³¹P (CDCl₃): 7.77 ppm ¹H (CDCl₃): 1.75–1.85 (m, 2H, CH₂S);1.90 (s, 3H, SCH₃); 2.01, 2.10 (s, 3H, 5CH₃); 2.30–2.50 (m, 2H, CH₂CH₂S); 3.45–3.60 (m, 1H CHNH); 3.94 (s, 2H, H5′); 5.05 (bs, 1H, H4′);5.90–6.00 (m, 1H, H2′); 6.40–6.50 (m, 1H, H3′); 6.93 (bs, 1H, H1′); 7.68(s, 1H, H6) ¹³C (CDCl₃): 11.91 (5CH₃); 14.46 (SCH₃); 29.58 (CH₃SCH₂CH₂);34.69 (SCH₂CH₂); 56.42 (CHNH); 65.07–65.13 (C5′); 86.39–86.52 (C4′);90.14 (C1′); 111.70 (C5); 125.48 (C2′); 134.77 (C3′); 138.91 (C6);152.61 (C2); 167.18 (C4); 180.84 (COOH) Mass: C₁₅H₂₂O₈N₃PS: 434 ((M−1),100); 435 ((M), 15) HPLC: RT=31.38 min

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(glycinyl phosphate) Cf 1163

Yield=75% ³¹P (CDCl₃): 11.72 ppm ¹H (CDCl₃): 1.83 (s, 3H, 5CH₃); 3.29(d, CH₂, J=7.9 Hz); 3.85–3.92 (m, 2H, H5′); 5.00 (s, 1H, H4′); 5.85–5.88(m, 1H, H2′); 6.38–6.41 (m, 1H, H3′), 6.88–6.90 (bs, 1H H1′); 7.54 (s,1H, H6) ¹³C (CDCl₃): 19.09 (5CH₃); 52.24 (CH₂); 72.74–72.81 (C5′);93.61–93.73 (C4′); 97.57 (C1′); 119.08 (C5); 132.80 (C2′); 141.89 (C3′);145.74 (C6); 159.87 (C2); 174.34 (C4); 186.03–186.15 (COOH) Mass:C₁₂H₁₆O₈N₃P: 360 ((M−1), 100); 361 ((M), 15) HPLC: RT=32.57 min

2′,3′-dideoxy-2′,3′-didehydrothymidino 5′-(phenylmethoxyisoleucinylphosphate) Cf 1186

Yield=82% ³¹P (CDCl₃): 4.59, 5.16 ppm ¹H (CDCl₃): 0.91–0.99 (m, 6H,CH₃+CH₃); 1.09–1.26 (CHCH₃); 1.28–1.56 (m, 2H, CH₂); 1.92, 1.97 (s, 3H,5CH₃); 3.60–3.77 (m, 1H, CHNH); 3.77 (s, 3H, OCH₃); 3.88–3.99 (m, 1H,NHCH); 4.30–4.52 (m, 2H, H5′); 5.11–5.13 (m, 1H, H4′); 5.95–6.00 (m, 1H,H2′); 6.35–6.45 (m, 1H, H3′); 7.10–7.13 (m, 1H, H1′); 7.16–7.45 (m, 6H,Ar+H6); 8.68 (bs, 1H, NH) ¹³C (CDCl₃); 11.90–11.92 (CH₂ CH₃);12.76–12.81 (5CH₃); 15.64 (CHCH₃); 25.06–25.14 (CH₂ CHCH₃);39.39–39.47–39.52–39.60 (CH₂); 52.61 (OCH₃); 59.38–59.54 (NHCH);66.94–67.58–67.65 (C5′); 84.91–85.04–85.16 (C4′); 89.94–90.21 (C1′);111.75–111.87 (C5′); 120–151 (m Ar); 127.82–127.87 (C2′); 133.49–133.69(C3′); 135.99–136.28 (C6); 151.37 (C2); 164.40 (C4);173.53–173.59–173.64 (COOCH₃) Mass: C₂₃H₃₀O₈N₃P: 529.91 ((M+Na)⁺, 100)HPLC: RT=30.52 and 31.14 min

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenylalaninyl phosphate) Cf1187

Yield=68% ³¹P (CDCl₃): 7.58 ppm ¹H (CDCl₃): 1.70 (s, 3H, 5CH₃);2.64–2.80 (m, 2H, CH ₂Ph); 3.57–3.64 (m, 1H, CHNH); 3.68–3.70 (m, 2H,H5′); 4.85 (s, 1H, H4′); 5.73–5.75 (m, 1H, H2′); 6.26–6.29 (m, 1H, H3′);6.74–6.75 (m, 1H, H1′); 7.02–7.28 (m, 5H, CH₂ Ph); 7.44 (s, 1H, H6) ¹³C(CDCl₃): 11.88 (5CH₃); 40.92–40.97 (CH₂ ala); 58.27 (CH ala);65.22–65.28 (C5′); 86.36–86.49 (C4′); 90.22 (C1′); 111.63 (C5); 125.38(C2′); 126–129 (m, Ar); 134.74 (C3′); 138.31–138.48 (C6); 152.40 (C2);166.81 (C4); 180.87–180.96 (COOH) Mass: C₁₉H₂₂O₈N₃P: 450 ((M−1)⁻, 100);451 ((M⁻, 20) HPLC: RT=32.11 min

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(valinyl phosphate) Cf 1190

Yield=67% ³¹P (CDCl₃): 8.35 ppm ¹H (CDCl₃): 0.72 (t, 6H, (CH ₃)₂CH,J=7.3 Hz); 1.62–1.73 (m, 1H, (CH₃)₂CH); 1.77 (s, 3H, 5CH₃); 3.12 (dd,1H, NHCH, J=5.6 Hz and 9.4 Hz); 3.80 (dd, 2H, H5′, J=3.5 Hz and 4.4 Hz);4.92 (s, 1H, H4′); 5.76–5.78 (m, 1H, H2′); 6.31–6.35 (m, 1H, H3′);6.79–6.81 (m, 1H, H1′); 7.53 (s, 1H, H6) ¹³C (CDCl₃): 11.84 (5CH₃);17.95–18.84 ((CH₃)₂CH); 32.30–32.38 ((CH₃)₂ CH); 62.43 (CHNH);65.18–65.24 (C5′); 86.43–86.58 (C4′); 90.25 (C1′); 111.65 (C5); 125.20(C2′); 134.90 (C3′); 138.73 (C6); 152.52 (C2); 167.05 (C4);181.27–181.31 (COOH) Mass: C₁₅H₂₂O₈N₃P: 402 ((M−1)⁻, 100); 403 ((M)⁻,30) HPLC: RT=31.90 min

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(leucinyl phosphate) Cf 1192

Yield=83% ³¹P (CDCl₃): 7.98 ppm ¹H (CDCl₃): 0.71 (d, 6H, (CH ₃)₂CH,J=6.5 Hz); 1.22–1.34 (m, 2H, CH₂); 1.34–1.71 (m, 1H, (CH₃)₂CH); 1.80 (s,3H, 5CH₃); 3.30–3.38 (m, 1H, CHNH); 3.82–3.85 (m, 2H, H5′); 4.95 (s, 1H,H4′); 5.80–5.82 (m, 1H, H2′); 6.35–6.37 (m, 1H, H3′); 6.81–6.82 (m, 1H,H1′); 7.58 (s, 1H, H6) ¹³C (CDCl₃): 12.53 (5CH₃); 22.88–22.99((CH₃)₂CH); 25.28 (CH₂); 45.27–45.34 ((CH₃)₂ CH); 56.38 (CHNH);65.74–65.81 (C5′); 87.12–87.25 (C4′); 90.89 (C1′); 112.30 (C5); 125.99(C2′); 135.49 (C3′); 139.44 (C6); 153.12 (C2); 167.70 (C4);183.36–183.42 (COOH) Mass: C₁₆H₂₄O₈N₃P: 416 ((M−1)⁻, 100); 417 ((M⁻, 20)HPLC: RT=35.02 min

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenylmethoxyalaninylphosphate) [fast diastereoisomer] Cf 1193

³¹P (CDCl₃): 4.51 ppm ¹H (CDCl₃) 1.25–1.40 (m, 3H, CHCH ₃); 1.86–1.90(m, 3H, 5CH₃); 3.74–3.90 (m, 4H, OCH₃+CH ala); 4.37–4.47 (m, 2H, H5′);5.08 (bs, 1H, H4′); 5.91–5.93 (m, 1H, H2′); 6.38–6.41 (m, 1H, H3′);7.07–7.09 (m, 1H, H1′); 7.20–7.39 (m, 6H, Ar+H6); 9.04 (bs, 1H, NH) ¹³C(CDCl₃): 10.85 (5CH₃); 19.38–19.45 (CHCH₃); 48.71 (CHCH₃); 51.14 (OCH₃);64.91–64.97 (C5′); 83.11–83.22 (C4′); 88.03 (C1′); 109.77 (C5); 118–149(m, Ar); 125.84 (C2′); 131.88 (C3′); 134.44 (C6); 149.34 (C2); 162.35(C4); 172.53–172.62 (CO ala)

2′,3′-dideoxy-2′,3′-didehydrothymidine 5′-(phenylprolinyl phosphate) Cf1194

Yield=41% ³¹P (CDCl₃): 5.27 ppm ¹H (CDCl₃): 1.55 (s, 3H, 5CH₃);1.56–2.15 (m, 4H, CHCH ₂CH ₂); 3.10–3.30 (m, 2H, NCH₂); 3.90–4.00 (m,1H, NCH); 4.20–4.50 (m, 2H, H5′); 5.11 (s, 1H, H4′); 5.89–5.91 (m, 1H,H2′); 6.41–6.44 (m, 1H, H3′); 6.76–6.78 (m, 1H, H1′); 6.99–7.40 (m, 6H,Ar+H6) ¹³C (CDCl₃): 11.84 (5CH₃); 25.44–25.56 (CH₂CH₂N); 31.94–32.06(CH₂CHN); 47.40–47.46 (NCH₂); 63.31 (CHN); 67.14–67.21 (C5′);85.56–85.68 (C4′); 90.69 (C1′); 111.00 (C5), 120–150 (m, Ar), 125.07(C2′), 134.13 (C3′), 138.26 (C6), 152.67 (C2), 166.64 (C4), 181.32(COOH) Mass: C₂₁H₂₄O₈N₃P: 476 ((M−1)⁻, 100); 477 ((M)⁻, 25) HPLC:RT=34.16 min

1001 2′,3′-dideoxy-2′,3′didehydroadenosine-5′-(phenyl methoxyalaninylphosphoramidate:

Yield=67% ¹H (dmso-d6): 8.14 (1H, s, H8), 8.06 (1H, d, H2), 7.07–7.40(7H, m, Phe-H & NH₂), 6.93 (1H, s, H1′), 6.47 (1H, 2d, H3′), 6.21 (1H,d, H3′), 5.96 (1H, m, NH), 5.11 (1H, m, H4′), 4.10 (2H, m, H5′),3.5–4.83 (1H, 2m, CH ala), 3.52 (3H, d, MeO), 1.08 (3H, 2d, CH₃ ala).³¹P (dmso-d6): 4.92, 4.78. ¹³C (dmso-d6): 172.909–172.815 (CO ala),154.663 (C-2), 152.238 (C-6), 149.524–149.442 (Ar-ipso), 148.782 (C-4),138.006–137.907 (C-8), 132.286–132.205 (C-2′), 128.621 (Ar-meta),125.384–125.210 (Ar para), 123.928 (C-3′), 119.067–119.00 (Ar ortho),118.508 (C-5), 87.311–87.060 (C-1′), 84.485–84.368 (C-4′), 66.093–65.324(C-5′), 51.477–51.429 (OMe), 49.109–48.989 (C—H ala), 19.903–19.585 (CH₃ala). Mass. Calculated MH⁺: 475.149. Found: 475.151

1093 2′,3′-dideoxy Adenosine 5′-(phenyl methoxyalaninyl) phosphoramidate

Yield=42% ¹H (CDCl₃): 8.32 (1H, s, H-8), 8.12 & 8.11 (1H, 2s, H-2) 7.22(5H, m, Ar), 6.40 (2H, 2bs, NH₂), 6.30 (1H, t, H-1′, J=5.4 Hz), 4.42(4H, m, N—H, 2H5′ & H4′), 4.00 (1H, 2d, Ala C—H), 3.65 (3H, 2s, OMe),2.52 (2H, m, H3′), 2.13 (2H, m, H2′), 1.31 (3H, 2d, CH₃ ala, J=7.3 Hz).³¹P (CDCl₃): 4.26, 4.19. ¹³C nmr (CDCl₃): 174.534, 174.468, 174.441,174.372 (O—C═O), 156.148 (C-2), 153.331 (C-6), 151.092 & 151.006 (2 Aripso), 149.674 & 149.599 (C-4), 139.211 & 139.103 (C-8), 130.040 (Armeta), 125.325 (Ar para), 120.570 (C-5), 120.508 & 120.327 (Ar ortho),85.994 & 85.746 (C-1′), 80.105, 79.985 & 79.874 (C-4′), 68.136, 68.067,67.704 & 67.636 (C-5′), 52.868 (OMe), 50.628 & 50.531 (Ala C—H), 32.712(C-2′), 26.339 & 26.106 (C-3′), 21.337, 21.264 & 21.190 (CH₃ ala). Mass:Calculated MH⁺: 477.165. Found: 477.164.

1094 2′,3′-dideoxy-2′,3′ didehydroadenosine 5′-(phenyl benzylalaninyl)phosphoramidate:

Yield=65% ¹H (CDCl₃): 8.32 (1H, bs, H-8), 7.99 (1H, bs, H-2), 7.21 (11H,m, Ar—H & H1′), 6.34 (1H, m, H3′), 6.07 (1H, m, H2′), 5.81 (2H, 2bs,NH₂), 5.08 (3H, 2bs, Bz-CH₂ & H4′), 4.05 (4H, m, NH, CH, H5′), 1.24 (3H,2d, methyl ala, J=6.9 Hz). ³¹P (CDCl₃): 4.21, 3.98 ¹³C (CDCl₃): 173.700& 173.601 (O—C═O), 156.005 (C-2), 153.728 (C-6), 150.952 & 150.870 (Ar),150.322 & 150.280 (C-4), 139.484 & 139.368 (C-8), 135.672 (Ar), 133.733& 133.654 (C-2′), 130.066 (Ar), 129.041, 128.895, 128.635 & 128.601(Ar), 126.751 & 126.598 (C-3′), 125.375 (Ar), 120.529, 120.463, 120.399,120.119 & 120.051 (C-5 & Ar), 88.702 & 88.476 (C-1′), 85.907, 85.476,85.791 & 85.736 (C-4′), 67.632, 67.475 & 67.403 (C-5′ and Bz-CH₂),66.805 & 66.745 (C-5′), 50.677 & 50.542 (Ala C—H), 21.399, 21.335,21.083 & 21.019 (methyl Ala). Mass: Calculated MH⁺: 551.181. Found:551.179.

1168 2′,3′-dideoxy-2′,3′-didehydroadenosine 5′-alaninyl phoshoramidate

Yield=69% ¹H nmr (D₂O): 8.09 (1H, s, H8), 7.88 (1H, s, H2), 6.81 (1H, s,H1′), 6.33 (1H, d, H3′), 6.02 (1H, d, H3′), 5.01 (1H, m, H4′), 4.73 (2H,m, H5′), 3.5–4.83 (1H, 2m, CH ala), 0.89 (3H, 2d, CH₃ ala). ³¹P (D₂O):8.34. ¹³C (D₂O): 183.055 (CO ala), 155.549 (C-2), 152.745 (C-6), 148.643(C-3), 140.928 (C-8), 134.730 (C-2′), 124.709 (C-3′), 118.527 (C-5),88.299 (C-1′), 87.199 & 87.073 (C-4′), 65.215–65.149 (C-5′), 52.564(Ala1 C—H), 21.435–21.381 (Ala CH₃).

1196—2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(phenyl dimethoxyglutaminyl phosphoramidate

Yield 33% ³¹P (CDCl₂) 4.14, 4.76 ¹H (CDCl₃) 1.81, 1.85 (5CH₃); 1.91–2.18(m, 2H, CH₂ Gln); 2.24–2.36 (m, 2H, CH₂ Gln); 3.64 (s, 3H, NMe); 3.69(s, 3H, OMe); 3.92–4.21 (m, 2H, H5′); 4.23–4.42 (m, 2H, CH Gln, NH Gln);5.00 (m, 1H, H4′); 5.91 (m, 1H, H2′); 6.31 (m, 1H, H3′); 7.01 (m, 1H,H1′), 7.03–7.34 (m, 6H, Ph, H6); 9.49 (s, 1H, NH) ¹³C (CDCl₃)12.32–12.36 (5CH₃); 29.01–29.42 (CH₂ Gln); 29.46 (NMe); 51.81 (CH Gln);52.65 (OMe); 53.65–53.92 (CH₂ Gln); 66.63–67.33 (C5′); 84.48–84.71(C4′); 89.57–89.83 (C1′); 111.29–111.44 (C5); 119.98–120.22 (Ph);125.21–125.26 (Ph); 127.39–127.50 (C2′); 129.74–129.78 (Ph);133.00–133.25 (C3′); 135.60–135.90 (C6); 150.98 (C2); 164.00–164.09(C4); 172.96–173.23 (CO, CON) Mass (ES): C₂₃H₂₉N₄O₉P: 536 (M⁺, 100); 537(MH⁺, 32)

1214—2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(phenyl dimethoxyasparaginyl) phosphoramidate

Yield 75% ³¹P (CDCl₃) 1.15, 2.20 ¹H (CDCl₃) 1.81, 1.86 (s, 3H, 5CH₃);2.49–2.92 (m, 2H, CH₂ Asn); 3.64 (s, 3H, NMe); 3.72 (s, 3H, OMe);4.04–4.26 (m, 2H, H5′); 4.28–4.43 (m, 2H, CH Asn, NH Asn); 5.05 (m, 1H,H4′); 5.89 (m, 1H, H2′); 6.31 (m, 1H, H3′); 7.01 (m, 1H, H1′); 7.14–7.33(m, 6H, Ph, H6); 8.46 (s, 1H, NH) ¹³C (CDCl₃) 12.28 (5CH₃); 51.01 (CHAsn); 52.09 (OMe); 52.94 (CH₂ Asn); 84.75 (C4′); 89.60 (C1′); 111.30(C5); 125–130 (Ph); 127.32–127.48 (C2′); 133.10–133.41 (C3′); 135.94(C6) Mass (ES): C₂₂H₂₇N₄O₉P: 522 (M⁺, 100); 523 (MH⁺, 31)

1215—2′,3′-Dideoxy-2′,3′-didehydrothymidine-5′-(phenylmethoxytryptophanyl) phosphoramidate

Yield 100% ³¹P (CDCl₃) 4.15, 4.57 ¹H (CDCl₃) 1.74 (s, 3H, 5CH₃); 3.16(m, 2H, CH₂ Trp); 3.60 (s, 3H, OMe); 3.75–4.05 (m, 2H, H5′); 4.10–4.33(m, 2H, CH Trp NH Trp); 4.84 (m, 1H, H4′); 5.79 (m, 1H, H2′); 6.15 (m,1H, H3′); 6.86 (m, 1H, H1′); 6.91 (m, 1H, H6); 7.00–7.49 (m, 10H, Ar);8.45 (s, 1H, NH Trp); 9.14 (s, 1H, NH) ³C (CDCl₃) 14.75 (5CH₃); 32.46(CH₂ Trp); 54.91 (CH Trp); 57.53–57.61 (OMe); 69 (C5′); 87.06 (C4′);92.03–92.25 (C1′); 111.63 (C5); 127.60 (C2′); 135.45–135.83 (C3′);138.11–138.62 (C6); 152.78–153.41 (C2); 166.28–166.40 (C4); 175.85 (CO)Mass (ES): C₂₈H₂₈N₄O₉P: 579 (M⁺, 100); 580 (M⁺, 43)

462 3′-Deoxy-3′-β-azidothymidine 5′-(phenyl methoxylalaninyl)phosphoramidate

¹H (CDCl₃): 1.39 (d, 3H, J=7.2 Hz, CH₃ ala), 1.94 (s, 3H 5-Me), 2.15 (d,1H, J=15.5 Hz, H2′), 2.68–2.79 (m, 1H, H2′), 3.72 (s, 3H, OMe),3.90–4.50 (m, 6H, H3′+H4′+H5′+NH+CHala), 6.18 (dd, 1H, J=7.5 and 3.1 Hz,H1′), 7.1–7.4 (m, 6H, Ph+H6), 8.82 (bs, 1H, NH). ¹³C (CDCl₃): 12.67(5-Me) -20.96, 21.29 (ala-Me), 38.50 (C2′), 50.16, 50.28 (CHala), 52.57(OMeala), 60.74 (C3′), 64.43 (C5′), 80.17 (C4′), 83.93 (C1′), 111.21(C5), 120.11 (Ar2), 125.18 (Ar4), 129.73 (Ar3), 135.18 (C6), 159.96(Ar1), 150.30 (C4), 163.49 (C2), 173.84 (COala). ³¹P (CDCl₃): 1.55 IR(CDCl₃): 3216, 2113, 1685 cm−1. Mass 509.1543 (MH⁺, 40%, calculated509.1549), 340(12), 250(17), 200(18). HPLC: RT=28.48 min.

536 3′-Deoxy-3′β-azidothymidine 5′-(m-trifluoromethylphenylmethoxylalaninyl) phosphoramidate

¹H (CDCl₃): 1.39, 1.40 (d, 3H, J=7.2 Hz, Me-ala), 1.92, 1.93 (s, 3H,5-CH₃), 2.15 (d, 1H, J=15.1 Hz, H2′), 2.71–2.80 (m, 1H, H2′), 3.70, 3.71(s, 3H, OMe), 3.90–4.50 (m, 6H, H3′+H4′+H5′+NH+CHala), 6.19 (dd, 1H,J=7.7 and 3.3 Hz, H1′), 7.41–7.46 (m, 5H, Ph+H6), 9.52 (bs, 1H, NH). ¹³C(CDCl₃): 12.58 (5-Me), 20.75, 20.83 (CH₃ ala), 38.33, 38.44 (C2′),50.15, 50.29 (CHala), 52.55 (OMeala) 60.77 (C3′), 64.72 (C5′), 80.05,80.35 (d, J=6.8 Hz, C4′), 83.94 (C1′), 111.25 (C5), 117.43 (Ar2),121.81, 121.86 (Ar4), 123.37 (q, J=273 Hz, CF₃), 123.74 (Ar6), 130.35(Ar5), 132.11 (q, J=33 Hz, Ar3), 135.11 (C6), 150.49 (C4), 150.62 (Ar1),163.78 (C2), 173.68, 173.87 (d, J=7.8 Hz, COala). ³¹P: 2.69 Mass 577(MH⁺, 40%) 340 (13), 268 (14), 250 (12). HPLC: RT=30.66 min.

550 3′-Deoxy-3′-β-azidothymidine 5′-(3,5-dichlorophenylmethoxylalaninyl) phosphoramidate

¹H (CDCl₃): 1.42 (d, 3H, J=6.8 Hz, Me-ala), 1.94, 1.95 (d, 3H, J=1.2 Hz,5-CH₃), 2.17, 2.18 (d, 1H, J=15.1 Hz, H2′), 2.76–2.85 (m, 1H, H2′),3.74, 3.75 (s, 3H, OMe), 3.90–4.50 (m, 6H, H3′+H4′+H5′+NH+CHala), 6.20(dd, 1H, J=7.7 and 3.3 Hz, H1′), 7.19 (m, 2H, Ar2) 7.27 (S, 1H, Ar4),7.41, 7.42 (s, 1H, H6), 9.04 (bs, 1H, NH). ¹³C: 12.65 (5-Me), 20.85,20.91 (CH₃ ala), 38.38, 38.48 (C2′), 50.18, 50.29 (CHala), 52.68(OMeala), 60.77 (C3′), 64.86, 64.93 (C5′), 79.80, 80.20 (d, J=8 Hz,C4′), 83.97 (C1′), 111.35 (C5), 117.28, 119.38 (d, J=6 Hz, Ar2), 125. 58(Ar4), 135.10 (C6), 135.46, 135.50 (Ar3), 145.35 (Ar1), 150.36 (C4),163.61 (C2), 173.64, 173.79 (COala). ³¹P: 2.83 Mass 577, 579, 581 (MHz⁺5:3:1:) 307, 309, 311 (12:8:2) 289 (10)

In Vitro Testing

Cells were infected with HIV-1 as previously described [Balzarini et al.AIDS (1991), 5, 21–28]. Briefly, 5×10⁵ cells per milliliter wereinfected with HIV-1 or HIV-2 at 100 CCID₅₀ (50% cell culture infectivedose) per milliliter of cell suspension. Then 100 μL of the infectedcell suspension was transferred to microtiter plate wells and mixed with100 μL of the appropriate dilutions of the test compounds. After 4 daysgiant cell formation was recorded microscopically in the HIV-infectedcell cultures [CEM], and after 5 days the number of viable cells wasdetermined by trypan blue staining of the HIV-infected cell cultures[MT4]. The 50% effective concentration (EC₅₀) and 50% cytoxicconcentration (CC₅₀) were defined as the compound concentrationsrequired to reduce by 50% the number of giant cells or viable cells inthe virus-infected and mock-infected cell cultures, respectively.

The anti-HIV-1 activities and toxicities of compounds were also assessedin two cell lines:

C8166 cells. Cells were grown in RPMI 1640 with 10% calf serum. 4×10⁴cells per microtiter plate well were mixed with 5-fold dilutions ofcompound prior to addition of 10 CCID₅₀ units of III-B strain of HIV-1and incubated for 5–7 days (Betbeder et al. Antiviral Chem. Chemother.1, 241–247, 1990). Formation of syncytia was examined from 2 dayspost-infection. Culture fluid was collected at 5–7 days and gp120antigen production measured by ELISA (Mahmood and Hay, J. Immunol.Meth., 151, 9–13, 1992). The EC₅₀ is that concentration of drug [in μM]required to reduce gp120 production by 50%. Cell viability of infectedand uninfected cells were assessed by the MTT-Formazen method (Pauwelset al. J. Virol. Meth. 20, 309–321, 1988).

JM cells JM cells, which are relatively resistant to the antiviraleffects of AZT and a number of its derivatives, were infected with HIV-1strains and the antiviral and toxic effects of compounds assessed as forC8166 cells. Both GB8 or IIIB strains of HIV1 were used, with nodelectable differences in the end-points noted.

Each assay was carried out in duplicate, on at least two separateoccasions, and data quoted are the average of each separate assay.

The compounds of the present invention have been shown to be activeagainst both HIV1 and HIV2 in both TK⁺ and TK⁻ cells as illustrated inTable 2.

TABLE 2 HIV1 in C8166/JM HIV2 in CEM TK⁻/CEM TK⁻ EC₅₀ EC₅₀ EC₅₀ EC₅₀Compound C8166 μM JM CEM TK⁻ μM CEM TK⁻ 730 0.0008 0.0008 0.016 0.06 d4T0.08 0.8 1.2 >100 (comparative)

As expected, d4T (comparative) loses activity in the kinase deficientcells (especially CEM TK⁻), whilst compound 730 of the invention retainsgood activity in both TK⁺ and TK⁻ against both HIV1 and HIV2. Compound730 of the invention is >1000 times more potent than d4T in TK⁻ cells.Surprisingly, the compound is 100-fold more potent than d4T in CEM TK⁻assays.

The potent activity of one compounds of the invention is furthersupported by the data in Table 3, which illustrates activity, toxicityand selectivity index of a series of compounds of the present invention.

The enhanced anti-viral potency and reduced cytotoxicity of thephosphate derivatives lead to very large improvements in The enhancedanti-viral potency and reduced cytotoxicity of the phosphate derivativeslead to very large improvements in selectivity index [defined asCC₅₀/EC₅₀] evidencing marked improvements in in vivo efficacy comparedto d4T (comparative).

Evidence that the compounds of the present invention are acting via apathway different to that of d4T or AZT is provided by the data of Table4.

As can be seen, whilst the potency of d4T (comparative) is much reducedin nucleoside resistant strains, the potency of the compounds of thepresent invention is largely maintained. Thus, it is clear that thecompounds of the present invention are not acting primarily via theconventional nucleoside 5′triphosphate derivative.

CEM and MT4 cells (at 400,000 cells/ml) and PBL cells (at 2,000,000cells/ml) were exposed to different concentrations of [³H] 324 andincubated at 37° C. for 24 hours. Then cells were washed twice with coldPBS and to the cell pellet was added 400 μl cold methanol 66%. Afterstanding on ice for 10 min, the cell extract was centrifuged and thesupernatent analayzed on HPLC. As shown in Table 5, intracellular D4T-MP(monophosphate) levels increased proportionally in function of theinitial concentration of 324 in all three cell lines tested. However,the increase of D4T-TP (triphosphate) levels slowed down at initial 324concentrations that were higher than 25 μM (for CEM and MT4 cells) orhigher than 1.0 μM (for PBL). Surprisingly, a metabolite (designated X)accumulated substantially and predominantly in all three cell types. Theaccumulation was proportional to the initial 324 concentration, and,again, was lower in PBL than CEM and MT4 cells.

When 1 mM 324 was incubated with high concentrations of hog liveresterase at 37° C. in Tris-HCl buffer containing 5 mM MgCl₂, atime-dependent formation of a metabolite was observed. This metabolitesco-eluted with the predominant metabolite (X) that was found in the cellextracts after incubation of the intact cells with [³H] 324. metaboliteX corresponds to a compound of formula (10), wherein Y is oxygen, X¹ isNH, X² is oxygen, B is thymine, R¹ is Me, R² is hydrogen.

Data on an expanded range of compounds is presented in Table 6 (d4Tanalogues) and Table 7 (dideoxy and 3′-β-substituted nucleosideanalogues) in which:

-   -   Cpd and Init: refer to the compound reference numbers;    -   Y: refers to the group:    -   Z: refers to the 3′-substituent on a deoxyribose sugar wherein        the substituent is in an “α” orientation (R⁹) unless designated        “up” which refers to a “β” orientation (R¹⁰);    -   B: refers to the heterocyclic nucleic acid base, present at C1′        in β-orientation; conventional one-letter base codes are used;        pyrimidine substituents are at C5.

The data columns are, in order:

-   HIV1 MT4: EC₅₀ in μM for inhibition of HIV-1 in MT4 cells.-   HIV2 MT4: EC₅₀ in μM for inhibition of HIV-2 in MT4 cells.-   CC50 MT4: CC₅₀ in μM for toxicity to MT-4 cells.-   HIV1 CEM: EC₅₀ in μM for inhibition of HIV-1 in CEM cells.-   HIV2 CEM: EC₅₀ in μM for inhibition of HIV-2 in CEM cells.-   HIV2 CEM-TK-: EC₅₀ in μM for inhibition of HIV-2 in CEM/TK⁻ cells.-   CC50 CEM: CC₅₀ in μM for toxicity to CEM cells.-   EC50 MSV: EC₅₀ in μM for inhibition of MSV-   MCC MSV: Minimum cytotoxic concentration in MSV assay

Where data of table 6 differs from that presented in Tables 2 to 5, thedata of the former relates to the mean result obtained from two or morerepeat experiments, whereas the latter relates to individualexperimental results.

TABLE 3 Selec- Tox- tivity Activity icity CC₅₀/EC₅₀ Entry Ar R¹ J EC₅₀CC₅₀ × 10³ 323 4-EtPh Me Me 0.0032 50 15.6 324 Ph Me Me 0.0032 150 46.9327 4-FPh Me Me 0.0032 200 62.5 526 3- Me Me 0.0008 200 250 CF₃Ph 5463,5- Me Me 0.001  100 100 Cl₂Ph 730 Ph Me Bzl 0.0008 400 500 776 2,4- MeMe 0.0008 100 125 Br₂Ph 779 F₅Ph Me Me 0.064  80 1.25 862 Ph Me Hex0.0012 500 417 863 Ph Me Me 0.016  500 31.2 864 Ph CH₂iPr Me0.016  >1000 >62.5 865 Ph iPr Me 0.8   >1000 >1.25 866 Ph H Me0.8   >1000 >1.25 867 Ph [CH₂]₂SMe Me 0.0016 >1000 >62.5 868 2,4- Me Bzl0.0032 500 156 Br₂Ph 877 Ph Bzl Bzl 0.0003 80 267 878 Ph Bzl tBu 0.16 150 0.9 892 Ph Me Cyclo- 0.0016 500 312 hex 893 Ph Me tBu0.2   >1000 >5.0 [data are μM for HIV1 in C8166 cells] By comparison,similar data for d4T: d4T — — — 0.08  50 0.6

TABLE 4 EC₅₀ EC₅₀ EC₅₀ SI EC₅₀ EC₅₀ HeLa HeLa EC₅₀ in μM 8166 8166 CEMCEM TK− HIV1 HIV1 Compound HIV RT HIV1 HIV1 HIV1 HIV2 d4T-Sensitived4T-Resistant d4T Inactive 0.08 625 0.5 >100 0.86 3.38 324 50 0.003262,500 0.18 0.08 n/d n/d 526 n/d 0.0008 >250,000 0.08 0.06 0.04 0.05 546n/d 0.001 >200,000 0.06 0.06 0.03 0.04 AZT Inactive 0.008 >100,0000.003 >100 n/d n/d

TABLE 5 Metabolism of [³H] 324 after 24 hr incubation in human CEM, MT4and PBL cells nmole/10⁹ cells CEM MT-4 PBL Initial concentration of [³H]324 (μM) Metabolite 0.2 1.0 5.0 25 100 500 0.2 25 0.2 1.0 25 324 + D4T7.6 47.8 228 897 4,333 16,691 7.9 1,255 2.0 12.2 245 D4T-MP 3.9 10.8 54490 2,259 11,359 29 394 2.4 14.2 355 D4T-DP 1.5 5.1 21.6 75 214 430 2.0116 0.45 1.8 15.3 D4T-TP 10.3 37.6 177 553 691 938 22.6 535 6.6 27 149133 628 3,164 16,193 66,359 204,442 117 14,582 17.6 97.3 1,995

TABLE 6 HIV1 HIV2 CC50 HIV1 HIV2 2 CEM CC50 EC50 MCC- Cpd Init ArO Y Z OM M M CEM CEM TK CEM MSV MSV 268 AS — — = T 0.24 1.2 >100 321 AS/ HexOHexO = T >42 >42 >42 36 DC 322 AS TECO MeValNH = T 29 71 59 323 AS EtPhOMeAlaNH = T 0.057 0.063 >100 0.07 0.16 0.06 60 324 AS/ PhO MeAlaNH = T0.081 0.053 >100 0.075 0.075 0.075 100 DC 325 AS TECO MePheNH = T 0.440.5 >100 1 2 0.7 326 AS PrO MeAlaNH = T 36 84 >250 >230 >230 135 ≧250345 AS TECO MeMetNH = T 8 11 10 400 AS TECO PntNA = T >40 >40 >40 401 ASTECO PrNH = T >210 >210 >210 402 AS TECO BuNH = T 118 >204 161 403 ASTECO EtNH = T >216 >216 >218 404 AS TECO PrO = T >209 >209 >209 406 ASTECO MeOC- = T >203 >203 >203 H2CH2 407 AS TECO HO = T 0.5 0.5 86 446 AS— — = U >95 >95 >95 479 SI iPrO iPrO = T >258 >258 >258 5′-PNH 180 SIBuO BuO = T >48 >48 >48 5′-PNMe 481 SI BuO BuO = T >9 >9 >9 5′-PNMe 504AS TFEO BuNH = T 73 116 ≧226 526 AS mCF3- MeAlaNH = T 0.05 0.11 10 0.150.15 0.12 30 PhO 546 AS 3,5- MeAlaNH = T 0.037 0.12 10.5 0.12 0.15 0.1226.9 Cl2PhO 547 AS mTFM- MeAlaNH = U >3520 >7 >35 PhO 551 AS EtO ElO =U >58 >58 >58 558 AS PhO MeAlaNH = U >44 >44 >44 561 AS FPhO MeAlaNH = U85 4 >72 562 AS TCEO TCEO = U >36 >36 >36 563 AS EtO PrNH =T >268 >268 >268 564 AS EtO MeAlaNH = T 28.5 62.65 ≧250 >48 >240 >48≧250 730 DC PhO BzAlaNH = T 0.016 0.016 0.06 25 740 DC MeO MeAlaNH = T25.4 50.9 >250 20 50 >250 >250 775 DC HO HO = T 0.8 0.95 33 174 776 Dc2,4- MeAlaNH = T 0.04 0.055 0.025 16 Br₂PhO 779 DC F5PO MeAlaNH = T 1.724.01 82 2.5 3.7 8.5 115 786 DC HexO H = T 0.6 0.5 30 150 787 DC MeO H =T 0.65 0.95 45 144 DigoIO 788 DC EtO H = T 0.65 0.6 30 115 789 DC DecO H= T 0.65 0.65 40 64 790 DC BuO H = T 0.65 0.9 30 177 791 DC OctO H = T0.4 0.6 23 90 792 DC PutO H = T 0.55 0.9 40 137 793 DC PtO H = T 0.650.65 33 170 816 DC Cl6O H = T 0.85 0.65 15 25 817 DC MeO H = T 0.7 0.5520 146 828 DC PhO Cl2NH = T >10 >10 14 829 DC PhO C8NH = T >10 ≧10 23840 DC PhO BuNH = T 140 100 125 >250 849 DC PhO BzAlaNH = U 23 15 10 115853 DC PhO OH = T 0.65 0.7 23 153 858 DC PhO PrNH =T >250 >250 >250 >250 859 DC PhO HxNH = T >50 >50 >50 145 860 DC PhOPolNH = T >250 >250 >250 ≧250 861 DC PhO CNEO = T 1.2 0.95 17.5 ≧250 862DC PhO HxAlaNH = T 0.06 0.055 0.033 48 863 DC PhO MePheNH = T 0.2 0.4 340.8 1.35 0.33 216 864 DC PhO MeLeuNH = T 0.2 0.5 62 1.1 2.23 0.4 ≧250865 DC PhO MeValNH = T 3.6 11.2 ≧250 12.5 12.5 4 >250 866 DC PhO MeGlyNH= T 1.3 6.7 ≧250 6 6 7 ≧250 867 DC PhO MeMetNH = T 0.2 0.4 22 0.6 0.80.34 ≧250 868 DC Br2- BzAlaNH = T 0.2 0.2 0.2 50 PhO 870 DC Br2- BzAlaNH= U >50 >50 12.5 180 PhO 877 DC PhO BzPheNH = T 0.6 0.6 0.24 44 878 DCPhO tBUPhe- = T 2 2 0.65 80 NH 879 DC PhO MeProNH = T >10 >10 >10 42.5880 DC PhO PhO = T 25 25 75 ≧250 881 DC HO NH2- = T AlaNH 892 GO/ PhOCHx- = T 0.065 0.075 0.09 51.4 D AlaNH 893 GO/ PhO tBuAla- = T 0.85 1.10.74 ≧250 D NH 932 ASS PhO Me D = T 3 2 2.5 >250 ≧100 >100 AlaNH 933 DCPhO BzProNH = T 35 12.5 >50 92 >100 >100 949 DC PhO EtMeNH = T 0.8 0.30.15 166 52 >100 950 DC PhO Et-β-AlaNH = T 250 ≧250 >250 >250 >100 >100951 DC PhO EtAlaNH = T 0.1 0.07 0.07 55 25 >100 978 DC PhO MetaclO = T40 50 >250 ≧250 >100 >100 979 DC PhO EtActO = T 28 23 160 >250 >100 >100980 DC PhO MeGlyco = T 27.5 50 >250 >250 >100 >100 981 DC PhO EtGlyco =T 12.5 12.5 150 ≧250 >100 >100 982 DC PhO MeMandO = T 1.7 0.65 15 9414 >100 983 DC M Me- hetero- = T ≧250 122 >250 >250 >100 >100 Eph- cycleedrin 1078 SV PhO Me2Asp- = T 0.55 0.65 0.33 209 31.4 >100 NH 1079 SV HOAspNH = T 1.6 2.5 70 ≧250 9.3 >100 1080 SV HO MeAsp- = T 3.5 5 110 ≧25030.3 >100 NH[SC] 1081 ASS PhO Me2Gl = T 8 5.33 16 ≧250 88.8 >100 uNH1083 ASS HO GluNH = T 8.5 5.5 >250 ≧250 54.6 >100 1095 ASS HO D-AlaNH =T 1.3 1.6 10 >250 0.42 >100 1129 SV HO MeAlaNH = T 2 4.5 50 >25047.4 >100 1131 SV HO OH = T 0.4 0.6 50 ≧250 6.7 >100 1133 LB PhO ElOGly-= T 75 87.5 >250 ≧250 >100 >100 NMe 1135 SV MeO BzAlaNH = T 10 1517.5 >250 >100 >100 1137 SV OH BzAlaNH = T 0.95 1.6 8 ≧250 15.7 >1001139 MW PhO OCNOH = T 15 15 >50 66.6 >20 >20 1156 LB HO MetNH = T 1.270.7 50 >250 16.2 >100 1163 LB HO GlyNH = T 2 5 130 >250 1186 LB PhOMelleNH = T 5 1187 LB HO PheNH = T 3.5 1189 YW PhO CHxCH2- = T 0.04AlaNH 1190 LB OH ValNH = T 0.7 1192 LB OH LeuNH = T 1.4 1193 LB PhOMeAlaNH = T [FAST ISOMER] 1194 LB PhO ProNH = T 6 1196 KT PhO MeGluta- =T 1.2 mlneNH 1197 HWT PhO Me β AlaNH = T >250 1198 HWT PhO MeGAD- =T >250 ANH 1199 HWT PhO MeCap- = T >250 roylNH 1200 HWT PhO MeOCOC- = T0.12 Me2Ala 1214 KT PhO Me- = T 0.6 Aspar- aglneN 1215 KT PhO MeTry- = T4 pNH 1216 HW OH β = T 0.7 AlaNH 1217 HWT OH Caproyl- = T 1.4 NH 1218 PSPhO PnAlaNH = T <0.08 1219 PS PhO neoPnt- = T <0.08 AlaNH 1220 PS PhOPhenethyl- = T 0.7 AlaNH 1224 HWT OH Me- = T 1 GABANH 1226 PS PhO1-Napth- = T <0.08 MethAla 1227 PS PhO 2-Napth- = T <0.08 MeAlaNH

TABLE 7 CC50 HIV1 HIV2 2.CEM CC50 EC50 MCC Cpd Init Aro Y Z B HIV1MHIV2M M CEM CEM TK CEM MSV MSV 462 PB PhO MeAlaNH N3-up T 33 11 121 27.540 30 499 PB — N3-up T 0.9 2.3 >250 3 4 >250 536 PB mCF3PhO MeAlaNHN3-up T 0.45 0.9 104 1 2 3 550 PB 3,5- MeAlaNH N3-up T 0.5 1 98 1.4 3 12Cl2PhO 569 PB — N3-up U >400 >400 >400 571 PB PhO MeAlaNH N3-upU >202 >202 117 657 ASS PhO HexNH N3-up T >40 >40 >18 659 ASS PhO BuNHN3-up T >42 >42 >42 661 ASS PhO C12NH N3-up T >7 >7 >7 687 DC — N3-upBzT 2.5 2.8 >100 >100 731 DC PhO BzAlaNH N3-up r 0.28 0.7 1.1 88 739 DCMeO MeAlaNH N3-up T 10 18 >250 >250 774 ASS PhO MeAlaNH N3-up N- >10 >1015 OctT 777 DC 2,4- MeAlaNH N3-up T 0.5 0.55 0.19 55 Br2PhO 780 DC FSPhOMeAlaNH N3-up T 23 33 100 106 846 ASS PhO CNEO N3-up T 13 14 >250 >250847 ASS TFEO CNEO N3-up T 12 9 >250 >250 850 ASS PhO OH N3-up I 189 >250 >250 855 ASS TFEO OH N3-up r 17.5 17.5 >250 >250 856 ASS HexOCNEO N3-up T 13 25 >250 >250 857 ASS HexO OH N3-up T 5 10 >250 >250 941ASS PhO MeO- N3-up T >50 >50 115 >100 >100 PheNH 1069 OW — — H A 4 817.5 >250 24.3 >100 1071 ASS HO HOC[0]- N3-up T 115 10250 >250 >250 >100 AlaNH 1093 OW PhO Me- H A 0.016 0.035 0.055 2.571.95 >20 AlaNH 1221 CY PhO Me- H C 0.6 AlaNH 1225 OW PhO Me- H T 1.2AlaNHIn Vivo TestingInhibitory Effects of Test Compounds on the Initiation of MSV-InducedTumour Formation in NRMI Mice and on the Survival of MSV InnoculatedNMRI Mice.

Mice infected with Moloney Sarcoma Virus [MSV] were treated daily witheither placebo, or d4T [at one of two doses] or with compound 324 at oneof the same [equi-molar] doses.

Two- to three-day old NMRI mice (weighing—2 gram) were innoculatedsubcutaneously (s.c.) in the left hind leg with 50 μl MSV (100 fociforming units, as measured by in vitro determination of thevirus-induced transformation of murine C3H embryo fibroblast cells). At4 to 5 days post-infection, tumours develop and rapidly increase involume upon further aging of the mice. Within 10 to 12 dayspost-infection, mice (then weighing—5 to 6 gram) die from the viralinfection. Drug treatment started 1 hour prior to infection of thevirus, and further compound administration was given daily i.p. for anadditional 3 days. The mean day of tumour initiation (±standarddeviation) and the mean day of survival of the mice (±standarddeviation) was calculated and statistical significance of the averagedelay of tumour formation and the mean day of survival in the treatedgroups versus the untreated (control) group was assessed by two-tailedstudent's t-test.

Whilst d4T failed to give any detectable delay in either tumourappearance or death, a significant effect on both parameters was seenwith high-dose compound 324, and an effect on the first diseaseparameter at low dose [FIG. 1].

1. A compound of the formula (1) wherein Ar is phenyl, naphthyl, orpyridyl, unsubstituted or substituted with from 1 to 3

substituents selected from the group consisting of halogeno, halomethyl,hydroxy, carboxy, carboxyalkyl, alkoxy, alkanoyl, alkanoyloxy, phenoxy,benzoyl, benzoyloxy, amino, alkylamino, dialkylamino, cyano, azido,nitro, mercapto, and alkylthio; Y is ═O or ═S; X¹ is —O—, —NR³—, —S—,—CR³R⁴—, —CR³W¹- or —CW¹W2-; each of R³ and R⁴ independently ishydrogen, alkyl, or phenyl, unsubstituted or substituted with from 1 to3 substituents selected from the group consisting of halogeno,halomethyl, hydroxy, carboxy, carboxyalkyl, alkoxy, alkanoyl,alkanoyloxy, phenoxy, benzoyl, benzoyloxy, amino, alkylamino,dialkylamino, cyano, azido, nitro, mercapto, and alkylthio; X⁶ is CH₂and X² is —O—; X³ is alkylene of 1 to 6 carbon atoms; —CR¹R²— in whicheach of R¹ and R² is hydrogen, alkyl of 1 to 6 carbon atoms, or phenyl;or when X³ is NH, and X⁴═Z═O, and J is other than H, CHR¹ in which R¹ is3-guanidinopropyl, mercaptomethyl, 2-amino-2-carboxyethyldithiomethyl,carboxymethyl, 2-carboxyethyl, carbamoylmethyl, 2-carbamoylethyl,1H-imidazolemethyl, 4-aminobutyl, 2-methylthioethyl, hydroxymethyl,1-hydroxyethyl, 1H-indol-3-ylmethyl, benzyl, or 4-hydroxybenzyl; X⁴ is═O or ═CH₂; X⁵ is a direct bond or is —CH₂—; Z is —O—, —NR⁵—, —S—,alkylene, or phenylene in which R5 is hydrogen, alkyl, or phenyl,unsubstituted or substituted with from 1 to 3 substituents selected fromthe group consisting of halogeno, halomethyl, hydroxy, carboxy,carboxyalkyl, alkoxy, alkanoyl, alkanoyloxy, phenoxy, benzoyl,benzoyloxy, amino, alkylamino, dialkylamino, cyano, azido, nitro,mercapto, and alkylthio; J is hydrogen, alkyl of 1 to 16 carbon atoms,cycloalkyl of 3 to 12 carbon atoms, benzyl, phenyl, unsubstituted orsubstituted with from 1 to 3 substituents selected from the groupconsisting of halogeno, halomethyl, hydroxy, carboxy, carboxyalkyl,alkoxy, alkanoyl, alkanoyloxy, phenoxy, benzoyl, benzoyloxy, amino,alkylamino, dialkylamino, cyano, azido, nitro, mercapto, and alkylthioor a heterocyclic group selected from the group consisting of pyrrolyl,imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, pyrrolidinyl,pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl,tetrahydrofuranyl, pyranyl, pyronyl, pyridyl, pyrazinyl, pyridazinyl,piperidyl, piperazinyl, morpholinyl, benzofuranyl, isobenzofuryl,indolyl, oxyindolyl, isoindolyl, indazolyl, indolinyl, 7-azaindolyl,isoindazolyl, benzopyranyl, coumarinyl, isocoumarinyl, quinolyl,isoquinolyl, napthridinyl, cinnolinyl, quinazolinyl, pyridopyridyl,benzoxazinyl, quinoxalinyl, chromenyl, chromanyl, isochromanyl, andcarbolinyl; Q is —O—, —NR⁶—, —S—, —CR⁶R⁷—, —CR⁶W³— or CW³W⁴—, each of R⁶and R⁷ independently is hydrogen, alkyl, or phenyl, unsubstituted orsubstituted with from 1 to 3 substituents selected from the groupconsisting of halogeno, halomethyl, hydroxy, carboxy, carboxyalkyl,alkoxy, alkanoyl, alkanoyloxy, phenoxy, benzoyl, benzoyloxy, amino,alkylamino, dialkylamino, cyano, azido, nitro, mercapto, and alkylthio;each of W¹, W², W³ and W⁴ independently is fluoro, chloro, bromo, oriodo; T¹ and T² are independently selected from hydrogen and CH₂R⁸,where R⁸ is selected from H, OH and F; B is a monovalent radical ofunsubstituted thymine, cytosine, adenine, or guanine, or a monovalentradical of substituted uracil, thymine, cytosine, adenine, or guanine inwhich the substituents are selected from halogeno, halomethyl, hydroxy,carboxyl, carboxylalkyl, alkoxy, alkanoyl, alkanoyloxy, phenoxy,benzoyl, benzoyloxy, amino, alkylamino, dialkylamino, cyano, azido,nitro, mercapto, alkylthio, alkyl, and benzyl; or a pharmaceuticallyacceptable salt or ester thereof.
 2. A compound according to claim 1wherein Y is oxygen; X¹ is NH; X³ is CHR¹; X⁴ is oxygen; and Z isoxygen; wherein R¹ is alkyl of 1 to 6 carbon atoms, phenyl or3-guanidinopropyl, mercaptomethyl, 2-amino-2-carboxyethyldithiomethyl,carboxymethyl, 2-carboxyethyl, carbamoylmethyl, 2-carbamoylethyl,1H-imidazolemethyl, 4-aminobutyl, 2-methylthioethyl, hydroxymethyl,1-hydroxyethyl, 1H-indol-3-ylmethyl, benzyl, or 4-hydroxybenzyl.
 3. Acompound of formula (10)

wherein Y is ═O or ═S; X¹ is —O—, —NR³—, —S—, —CR³R⁴—, —CR³W¹— or—CW¹W²—; each of R³ and R⁴ independently is hydrogen, alkyl, or phenyl,unsubstituted or substituted with from 1 to 3 substituents selected fromthe group consisting of halogeno, halomethyl, hydroxy, carboxy,carboxyalkyl, alkoxy, alkanoyl, alkanoyloxy, phenoxy, benzoyl,benzoyloxy, amino, alkylamino, dialkylamino, cyano, azido, nitro,mercapto, and alkylthio; X⁶ CH₂ and X² is —O—; X³ is alkylene of 1 to 6carbon atoms; —CR¹R²— in which each of R¹ and R² is hydrogen, alkyl of 1to 6 carbon atoms, or phenyl; or CHR¹ in which R¹ is 3-guanidinopropyl,mercaptomethyl, 2-amino-2-carboxyethyldithiomethyl, carboxymethyl,2-carboxyethyl, carbamoylmethyl, 2-carbamoylethyl, 1H-imidazolemethyl,4-aminobutyl, 2-methylthioethyl, hydroxymethyl, 1-hydroxymethyl,1H-indol-3-ylmethyl, benzyl, or 4-hydroxybenzyl; X⁴is ═O or ═CH₂; X⁵ isa direct bond or is CH₂—; Q is —O—, —NR⁶—, —S—, —CR⁶R⁷—, —CR⁶W³— or—CW³W⁴—, each of R⁶ and R⁷ independently is hydrogen, alkyl, or phenyl,unsubstituted or substituted with from 1 to 3 substituents selected fromthe group consisting of halogeno, halomethyl, hydroxy, carboxy,carboxyalkyl, alkoxy, alkanoyl, alkanoyloxy, phenoxy, benzoyl,benzoyloxy, amino, alkylamino, dialkylamino, cyano, azido, nitro,mercapto, and alkylthio; each of W¹, W², W³ and W⁴ independently isfluoro, chloro, bromo, or iodo; T¹ and T² are independently selectedfrom hydrogen and CH₂R⁸, where R⁸ is selected from H, OH and F; B is amonovalent radical of unsubstituted thymine, cytosine, adenine, orguanine, or a monovalent radical of substituted uracil, thymine,cytosine, adenine, or guanine in which the substituents are selectedfrom halogeno, halomethyl, hydroxy, carboxyl, carboxylalkyl, alkoxy,alkanoyl, alkanoyloxy, phenoxy, benzoyl, benzoyloxy, amino, alkylamino,dialkylamino, cyano, azido, nitro, mercapto, alkylthio, alkyl, andbenzyl; or pharmaceutically acceptable salt or ester thereof.
 4. Acompound according to claim 3 wherein Y is oxygen; X¹ is NH; X³ is CHR¹;X⁴ is oxygen; wherein R¹ is alkyl of 1 to 6 carbon atoms, phenyl or3-guanidinopropyl, mercaptomethyl, 2-amino-2-carboxyethyldithiomethyl,carboxymethyl, 2-carboxyethyl, carbamoylmethyl, 2-carbamoylethyl,1H-imidazolemethyl, 4-aminobutyl, 2-methylthioethyl, hydroxymethyl,1-hydroxyethyl, 1H-indol-3-ylmethyl, benzyl, or 4-hydroxybenzyl.
 5. Acompound according to claim 1 wherein B is adenine or thymine.
 6. Acompound according to claim 5 wherein B is adenine.
 7. A method oftreatment of HIV infection comprising administration to a patient inneed of such treatment an effective dose of a compound according toclaim
 1. 8. A pharmaceutical composition comprising a compound accordingto claim 1 in combination with a pharmaceutically acceptable excipient.